Substances used for the detection, identification, analysis, etc. of chemical, biological, or pathologic processes or conditions. Indicators are substances that change in physical appearance, e.g., color, at or approaching the endpoint of a chemical titration, e.g., on the passage between acidity and alkalinity. Reagents are substances used for the detection or determination of another substance by chemical or microscopical means, especially analysis. Types of reagents are precipitants, solvents, oxidizers, reducers, fluxes, and colorimetric reagents. (From Grant & Hackh's Chemical Dictionary, 5th ed, p301, p499)
Chemical agents that react with SH groups. This is a chemically diverse group that is used for a variety of purposes. Among these are enzyme inhibition, enzyme reactivation or protection, and labelling.
International collective of humanitarian organizations led by volunteers and guided by its Congressional Charter and the Fundamental Principles of the International Red Cross Movement, to provide relief to victims of disaster and help people prevent, prepare for, and respond to emergencies.
Reagents with two reactive groups, usually at opposite ends of the molecule, that are capable of reacting with and thereby forming bridges between side chains of amino acids in proteins; the locations of naturally reactive areas within proteins can thereby be identified; may also be used for other macromolecules, like glycoproteins, nucleic acids, or other.
Narrow pieces of material impregnated or covered with a substance used to produce a chemical reaction. The strips are used in detecting, measuring, producing, etc., other substances. (From Dorland, 28th ed)
Serological reactions in which an antiserum against one antigen reacts with a non-identical but closely related antigen.
Commercially prepared reagent sets, with accessory devices, containing all of the major components and literature necessary to perform one or more designated diagnostic tests or procedures. They may be for laboratory or personal use.
Prepaid health and hospital insurance plan.
Studies in which the presence or absence of disease or other health-related variables are determined in each member of the study population or in a representative sample at one particular time. This contrasts with LONGITUDINAL STUDIES which are followed over a period of time.
Deliberate breeding of two different individuals that results in offspring that carry part of the genetic material of each parent. The parent organisms must be genetically compatible and may be from different varieties or closely related species.
Compounds containing the -SH radical.
A sulfhydryl reagent that is widely used in experimental biochemical studies.
A thiol-containing non-essential amino acid that is oxidized to form CYSTINE.
The rate dynamics in chemical or physical systems.
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
A standard reagent for the determination of reactive sulfhydryl groups by absorbance measurements. It is used primarily for the determination of sulfhydryl and disulfide groups in proteins. The color produced is due to the formation of a thio anion, 3-carboxyl-4-nitrothiophenolate.
A subclass of IMIDES with the general structure of pyrrolidinedione. They are prepared by the distillation of ammonium succinate. They are sweet-tasting compounds that are used as chemical intermediates and plant growth stimulants.
Organic salts or esters of methanesulfonic acid.
The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.
Method of analyzing chemicals using automation.
Analogs of those substrates or compounds which bind naturally at the active sites of proteins, enzymes, antibodies, steroids, or physiological receptors. These analogs form a stable covalent bond at the binding site, thereby acting as inhibitors of the proteins or steroids.
A reagent that is highly selective for the modification of arginyl residues. It is used to selectively inhibit various enzymes and acts as an energy transfer inhibitor in photophosphorylation.
The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH = log 1/2[1/(H+)], where (H+) is the hydrogen ion concentration in gram equivalents per liter of solution. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
Antibodies produced by a single clone of cells.
Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed)
The parts of a macromolecule that directly participate in its specific combination with another molecule.
Genetic loci associated with a QUANTITATIVE TRAIT.
The location of the atoms, groups or ions relative to one another in a molecule, as well as the number, type and location of covalent bonds.
A cytotoxic sulfhydryl reagent that inhibits several subcellular metabolic systems and is used as a tool in cellular physiology.
The phenomenon whereby compounds whose molecules have the same number and kind of atoms and the same atomic arrangement, but differ in their spatial relationships. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 5th ed)
The simultaneous or sequential binding of multiple cell surface receptors to different ligands resulting in coordinated stimulation or suppression of signal transduction.
Esters of the hypothetical imidic acids. They react with amines or amino acids to form amidines and are therefore used to modify protein structures and as cross-linking agents.
Any method used for determining the location of and relative distances between genes on a chromosome.
Elements of limited time intervals, contributing to particular results or situations.
Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.
Any technique by which an unknown color is evaluated in terms of standard colors. The technique may be visual, photoelectric, or indirect by means of spectrophotometry. It is used in chemistry and physics. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results.
Organic or inorganic compounds that contain the -N3 group.
Preservative for wines, soft drinks, and fruit juices and a gentle esterifying agent.
A reagent commonly used in biochemical studies as a protective agent to prevent the oxidation of SH (thiol) groups and for reducing disulphides to dithiols.
The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments.
A basic science concerned with the composition, structure, and properties of matter; and the reactions that occur between substances and the associated energy exchange.
Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.
Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.
A technique using antibodies for identifying or quantifying a substance. Usually the substance being studied serves as antigen both in antibody production and in measurement of antibody by the test substance.
A series of steps taken in order to conduct research.
The sum of the weight of all the atoms in a molecule.
Chloride and mercury-containing derivatives of benzoic acid.
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
The composition, conformation, and properties of atoms and molecules, and their reaction and interaction processes.
A system for verifying and maintaining a desired level of quality in a product or process by careful planning, use of proper equipment, continued inspection, and corrective action as required. (Random House Unabridged Dictionary, 2d ed)
A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.
A chemical reaction in which an electron is transferred from one molecule to another. The electron-donating molecule is the reducing agent or reductant; the electron-accepting molecule is the oxidizing agent or oxidant. Reducing and oxidizing agents function as conjugate reductant-oxidant pairs or redox pairs (Lehninger, Principles of Biochemistry, 1982, p471).
The genetic process of crossbreeding between genetically dissimilar parents to produce a hybrid.
Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are linear polypeptides that are normally synthesized on RIBOSOMES.
An organic mercurial used as a sulfhydryl reagent.
Studies determining the effectiveness or value of processes, personnel, and equipment, or the material on conducting such studies. For drugs and devices, CLINICAL TRIALS AS TOPIC; DRUG EVALUATION; and DRUG EVALUATION, PRECLINICAL are available.
The species Oryctolagus cuniculus, in the family Leporidae, order LAGOMORPHA. Rabbits are born in burrows, furless, and with eyes and ears closed. In contrast with HARES, rabbits have 22 chromosome pairs.
Agents that emit light after excitation by light. The wave length of the emitted light is usually longer than that of the incident light. Fluorochromes are substances that cause fluorescence in other substances, i.e., dyes used to mark or label other compounds with fluorescent tags.
An alkylating sulfhydryl reagent. Its actions are similar to those of iodoacetate.
Established cell cultures that have the potential to propagate indefinitely.
The covalent bonding of an alkyl group to an organic compound. It can occur by a simple addition reaction or by substitution of another functional group.
Organic compounds containing a carbonyl group in the form -CHO.
The restriction of a characteristic behavior, anatomical structure or physical system, such as immune response; metabolic response, or gene or gene variant to the members of one species. It refers to that property which differentiates one species from another but it is also used for phylogenetic levels higher or lower than the species.
The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain).
The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms.
The art or process of comparing photometrically the relative intensities of the light in different parts of the spectrum.
The development and use of techniques and equipment to study or perform chemical reactions, with small quantities of materials, frequently less than a milligram or a milliliter.
An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed.
The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.
A basis of value established for the measure of quantity, weight, extent or quality, e.g. weight standards, standard solutions, methods, techniques, and procedures used in diagnosis and therapy.
Test for tissue antigen using either a direct method, by conjugation of antibody with fluorescent dye (FLUORESCENT ANTIBODY TECHNIQUE, DIRECT) or an indirect method, by formation of antigen-antibody complex which is then labeled with fluorescein-conjugated anti-immunoglobulin antibody (FLUORESCENT ANTIBODY TECHNIQUE, INDIRECT). The tissue is then examined by fluorescence microscopy.
The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.
Organic compounds that generally contain an amino (-NH2) and a carboxyl (-COOH) group. Twenty alpha-amino acids are the subunits which are polymerized to form proteins.
Hydroxylated benzoic acid derivatives that contain mercury. Some of these are used as sulfhydryl reagents in biochemical studies.
The methyl imidoester of suberic acid used to produce cross links in proteins. Each end of the imidoester will react with an amino group in the protein molecule to form an amidine.
Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.
The facilitation of a chemical reaction by material (catalyst) that is not consumed by the reaction.
The property of antibodies which enables them to react with some ANTIGENIC DETERMINANTS and not with others. Specificity is dependent on chemical composition, physical forces, and molecular structure at the binding site.
Protection conferred on a host by inoculation with one strain or component of a microorganism that prevents infection when later challenged with a similar strain. Most commonly the microorganism is a virus.
Iodinated derivatives of acetic acid. Iodoacetates are commonly used as alkylating sulfhydryl reagents and enzyme inhibitors in biochemical research.
A chemical element having an atomic weight of 106.4, atomic number of 46, and the symbol Pd. It is a white, ductile metal resembling platinum, and following it in abundance and importance of applications. It is used in dentistry in the form of gold, silver, and copper alloys.
A water-soluble, enzyme co-factor present in minute amounts in every living cell. It occurs mainly bound to proteins or polypeptides and is abundant in liver, kidney, pancreas, yeast, and milk.
Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.
Chemical groups containing the covalent disulfide bonds -S-S-. The sulfur atoms can be bound to inorganic or organic moieties.
A toxic thiol mercury salt formerly used as a diuretic. It inhibits various biochemical functions, especially in mitochondria, and is used to study those functions.
The total number of cases of a given disease in a specified population at a designated time. It is differentiated from INCIDENCE, which refers to the number of new cases in the population at a given time.
A class of compounds of the type R-M, where a C atom is joined directly to any other element except H, C, N, O, F, Cl, Br, I, or At. (Grant & Hackh's Chemical Dictionary, 5th ed)
A group of compounds derived from ammonia by substituting organic radicals for the hydrogens. (From Grant & Hackh's Chemical Dictionary, 5th ed)
Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.
Serum that contains antibodies. It is obtained from an animal that has been immunized either by ANTIGEN injection or infection with microorganisms containing the antigen.
Methods utilizing the principles of MICROFLUIDICS for sample handling, reagent mixing, and separation and detection of specific components in fluids.
Organic mercury compounds in which the mercury is attached to a phenyl group. Often used as fungicides and seed treatment agents.
An analytical method used in determining the identity of a chemical based on its mass using mass analyzers/mass spectrometers.
The marking of biological material with a dye or other reagent for the purpose of identifying and quantitating components of tissues, cells or their extracts.
Specialized non-fenestrated tightly-joined ENDOTHELIAL CELLS with TIGHT JUNCTIONS that form a transport barrier for certain substances between the cerebral capillaries and the BRAIN tissue.
Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA).
An antineoplastic agent with alkylating properties. It also acts as a mutagen by damaging DNA and is used experimentally for that effect.
Spectroscopic method of measuring the magnetic moment of elementary particles such as atomic nuclei, protons or electrons. It is employed in clinical applications such as NMR Tomography (MAGNETIC RESONANCE IMAGING).
The relationship between the chemical structure of a compound and its biological or pharmacological activity. Compounds are often classed together because they have structural characteristics in common including shape, size, stereochemical arrangement, and distribution of functional groups.
Partial proteins formed by partial hydrolysis of complete proteins or generated through PROTEIN ENGINEERING techniques.
Proteins prepared by recombinant DNA technology.
In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.
Immunoglobulin molecules having a specific amino acid sequence by virtue of which they interact only with the ANTIGEN (or a very similar shape) that induced their synthesis in cells of the lymphoid series (especially PLASMA CELLS).
Linear POLYPEPTIDES that are synthesized on RIBOSOMES and may be further modified, crosslinked, cleaved, or assembled into complex proteins with several subunits. The specific sequence of AMINO ACIDS determines the shape the polypeptide will take, during PROTEIN FOLDING, and the function of the protein.
Theoretical representations that simulate the behavior or activity of biological processes or diseases. For disease models in living animals, DISEASE MODELS, ANIMAL is available. Biological models include the use of mathematical equations, computers, and other electronic equipment.
Classic quantitative assay for detection of antigen-antibody reactions using a radioactively labeled substance (radioligand) either directly or indirectly to measure the binding of the unlabeled substance to a specific antibody or other receptor system. Non-immunogenic substances (e.g., haptens) can be measured if coupled to larger carrier proteins (e.g., bovine gamma-globulin or human serum albumin) capable of inducing antibody formation.
The analysis of a chemical substance by inserting a sample into a carrier stream of reagent using a sample injection valve that propels the sample downstream where mixing occurs in a coiled tube, then passes into a flow-through detector and a recorder or other data handling device.
A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).
Determination, by measurement or comparison with a standard, of the correct value of each scale reading on a meter or other measuring instrument; or determination of the settings of a control device that correspond to particular values of voltage, current, frequency or other output.
Hydrocarbons with at least one triple bond in the linear portion, of the general formula Cn-H2n-2.
A phenotypically recognizable genetic trait which can be used to identify a genetic locus, a linkage group, or a recombination event.
The movement of materials (including biochemical substances and drugs) through a biological system at the cellular level. The transport can be across cell membranes and epithelial layers. It also can occur within intracellular compartments and extracellular compartments.
Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.
Techniques for labeling a substance with a stable or radioactive isotope. It is not used for articles involving labeled substances unless the methods of labeling are substantively discussed. Tracers that may be labeled include chemical substances, cells, or microorganisms.
A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.
Method of tissue preparation in which the tissue specimen is frozen and then dehydrated at low temperature in a high vacuum. This method is also used for dehydrating pharmaceutical and food products.
The circulation in a portion of the body of one individual of blood supplied from another individual.
The adaptive superiority of the heterozygous GENOTYPE with respect to one or more characters in comparison with the corresponding HOMOZYGOTE.
The intracellular transfer of information (biological activation/inhibition) through a signal pathway. In each signal transduction system, an activation/inhibition signal from a biologically active molecule (hormone, neurotransmitter) is mediated via the coupling of a receptor/enzyme to a second messenger system or to an ion channel. Signal transduction plays an important role in activating cellular functions, cell differentiation, and cell proliferation. Examples of signal transduction systems are the GAMMA-AMINOBUTYRIC ACID-postsynaptic receptor-calcium ion channel system, the receptor-mediated T-cell activation pathway, and the receptor-mediated activation of phospholipases. Those coupled to membrane depolarization or intracellular release of calcium include the receptor-mediated activation of cytotoxic functions in granulocytes and the synaptic potentiation of protein kinase activation. Some signal transduction pathways may be part of larger signal transduction pathways; for example, protein kinase activation is part of the platelet activation signal pathway.
The major immunoglobulin isotype class in normal human serum. There are several isotype subclasses of IgG, for example, IgG1, IgG2A, and IgG2B.
Sites on an antigen that interact with specific antibodies.
The genetic constitution of the individual, comprising the ALLELES present at each GENETIC LOCUS.
Proteins found in any species of bacterium.
Materials that add an electron to an element or compound, that is, decrease the positiveness of its valence. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 5th ed)
Carrier of aroma of butter, vinegar, coffee, and other foods.
A derivative of ACETIC ACID that contains one IODINE atom attached to its methyl group.
Alkyl compounds containing a hydroxyl group. They are classified according to relation of the carbon atom: primary alcohols, R-CH2OH; secondary alcohols, R2-CHOH; tertiary alcohols, R3-COH. (From Grant & Hackh's Chemical Dictionary, 5th ed)
Tests that are dependent on the clumping of cells, microorganisms, or particles when mixed with specific antiserum. (From Stedman, 26th ed)
Measurement of the intensity and quality of fluorescence.
Incorporation of biotinyl groups into molecules.
Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.
Determination of the spectra of ultraviolet absorption by specific molecules in gases or liquids, for example Cl2, SO2, NO2, CS2, ozone, mercury vapor, and various unsaturated compounds. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
Mercury-containing benzoic acid derivatives.
Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen.
A very strong halogenated derivative of acetic acid. It is used in acid catalyzed reactions, especially those where an ester is cleaved in peptide synthesis.
Passive agglutination tests in which antigen is adsorbed onto latex particles which then clump in the presence of antibody specific for the adsorbed antigen. (From Stedman, 26th ed)
Changing an open-chain hydrocarbon to a closed ring. (McGraw-Hill Dictionary of Scientific and Technical Terms, 5th ed)
A mass spectrometry technique used for analysis of nonvolatile compounds such as proteins and macromolecules. The technique involves preparing electrically charged droplets from analyte molecules dissolved in solvent. The electrically charged droplets enter a vacuum chamber where the solvent is evaporated. Evaporation of solvent reduces the droplet size, thereby increasing the coulombic repulsion within the droplet. As the charged droplets get smaller, the excess charge within them causes them to disintegrate and release analyte molecules. The volatilized analyte molecules are then analyzed by mass spectrometry.
A serine endopeptidase that is formed from TRYPSINOGEN in the pancreas. It is converted into its active form by ENTEROPEPTIDASE in the small intestine. It catalyzes hydrolysis of the carboxyl group of either arginine or lysine. EC
Theoretical representations that simulate the behavior or activity of chemical processes or phenomena; includes the use of mathematical equations, computers, and other electronic equipment.
Unsaturated hydrocarbons of the type Cn-H2n, indicated by the suffix -ene. (Grant & Hackh's Chemical Dictionary, 5th ed, p408)
Organic compounds which contain mercury as an integral part of the molecule.
A chelating agent that sequesters a variety of polyvalent cations such as CALCIUM. It is used in pharmaceutical manufacturing and as a food additive.
Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion.
Inorganic or organic salts and esters of boric acid.
Chromatography on non-ionic gels without regard to the mechanism of solute discrimination.
An atom or group of atoms that have a positive or negative electric charge due to a gain (negative charge) or loss (positive charge) of one or more electrons. Atoms with a positive charge are known as CATIONS; those with a negative charge are ANIONS.
The co-inheritance of two or more non-allelic GENES due to their being located more or less closely on the same CHROMOSOME.
Red blood cells. Mature erythrocytes are non-nucleated, biconcave disks containing HEMOGLOBIN whose function is to transport OXYGEN.
Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes.
Immunologic techniques based on the use of: (1) enzyme-antibody conjugates; (2) enzyme-antigen conjugates; (3) antienzyme antibody followed by its homologous enzyme; or (4) enzyme-antienzyme complexes. These are used histologically for visualizing or labeling tissue specimens.
Organic compounds that contain the (-NH2OH) radical.
The study of the structure, preparation, properties, and reactions of carbon compounds. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.
Bifunctional cross-linking agent that links covalently free amino groups of proteins or polypeptides, including those in cell membranes. It is used as reagent or fixative in immunohistochemistry and is a proposed antisickling agent.
Aminobenzenesulfonic acids. Organic acids that are used in the manufacture of dyes and organic chemicals and as reagents.
2,2-Dihydroxy-1H-indene-1,3-(2H)-dione. Reagent toxic to skin and mucus membranes. It is used in chemical assay for peptide bonds, i.e., protein determinations and has radiosensitizing properties.
Covalent attachment of HALOGENS to other compounds.
Predetermined sets of questions used to collect data - clinical data, social status, occupational group, etc. The term is often applied to a self-completed survey instrument.
The level of protein structure in which combinations of secondary protein structures (alpha helices, beta sheets, loop regions, and motifs) pack together to form folded shapes called domains. Disulfide bridges between cysteines in two different parts of the polypeptide chain along with other interactions between the chains play a role in the formation and stabilization of tertiary structure. Small proteins usually consist of only one domain but larger proteins may contain a number of domains connected by segments of polypeptide chain which lack regular secondary structure.
A 60-kDa extracellular protein of Streptomyces avidinii with four high-affinity biotin binding sites. Unlike AVIDIN, streptavidin has a near neutral isoelectric point and is free of carbohydrate side chains.
Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake.
A molecule that binds to another molecule, used especially to refer to a small molecule that binds specifically to a larger molecule, e.g., an antigen binding to an antibody, a hormone or neurotransmitter binding to a receptor, or a substrate or allosteric effector binding to an enzyme. Ligands are also molecules that donate or accept a pair of electrons to form a coordinate covalent bond with the central metal atom of a coordination complex. (From Dorland, 27th ed)
The status during which female mammals carry their developing young (EMBRYOS or FETUSES) in utero before birth, beginning from FERTILIZATION to BIRTH.
An essential amino acid. It is often added to animal feed.
Chemicals that bind to and remove ions from solutions. Many chelating agents function through the formation of COORDINATION COMPLEXES with METALS.
The mating of plants or non-human animals which are closely related genetically.
A characteristic showing quantitative inheritance such as SKIN PIGMENTATION in humans. (From A Dictionary of Genetics, 4th ed)
A technology, in which sets of reactions for solution or solid-phase synthesis, is used to create molecular libraries for analysis of compounds on a large scale.
Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. Primers are used extensively in genetic and molecular biology techniques.
An essential amino acid that is required for the production of HISTAMINE.
Complex nucleoprotein structures which contain the genomic DNA and are part of the CELL NUCLEUS of PLANTS.
An aspect of personal behavior or lifestyle, environmental exposure, or inborn or inherited characteristic, which, on the basis of epidemiologic evidence, is known to be associated with a health-related condition considered important to prevent.
A brominating agent that replaces hydrogen atoms in benzylic or allylic positions. It is used in the oxidation of secondary alcohols to ketones and in controlled low-energy brominations. (From Miall's Dictionary of Chemistry, 5th ed; Hawley's Condensed Chemical Dictionary, 12th ed,).
The relationship between the dose of an administered drug and the response of the organism to the drug.
The property of emitting radiation while being irradiated. The radiation emitted is usually of longer wavelength than that incident or absorbed, e.g., a substance can be irradiated with invisible radiation and emit visible light. X-ray fluorescence is used in diagnosis.
The vapor state of matter; nonelastic fluids in which the molecules are in free movement and their mean positions far apart. Gases tend to expand indefinitely, to diffuse and mix readily with other gases, to have definite relations of volume, temperature, and pressure, and to condense or liquefy at low temperatures or under sufficient pressure. (Grant & Hackh's Chemical Dictionary, 5th ed)
Compounds used extensively as acetylation, oxidation and dehydrating agents and in the modification of proteins and enzymes.
A reagent that forms fluorescent conjugation products with primary amines. It is used for the detection of many biogenic amines, peptides, and proteins in nanogram quantities in body fluids.
An enzyme of the oxidoreductase class that catalyzes the conversion of beta-D-glucose and oxygen to D-glucono-1,5-lactone and peroxide. It is a flavoprotein, highly specific for beta-D-glucose. The enzyme is produced by Penicillium notatum and other fungi and has antibacterial activity in the presence of glucose and oxygen. It is used to estimate glucose concentration in blood or urine samples through the formation of colored dyes by the hydrogen peroxide produced in the reaction. (From Enzyme Nomenclature, 1992) EC
A procedure consisting of a sequence of algebraic formulas and/or logical steps to calculate or determine a given task.
A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.
Procedures for finding the mathematical function which best describes the relationship between a dependent variable and one or more independent variables. In linear regression (see LINEAR MODELS) the relationship is constrained to be a straight line and LEAST-SQUARES ANALYSIS is used to determine the best fit. In logistic regression (see LOGISTIC MODELS) the dependent variable is qualitative rather than continuously variable and LIKELIHOOD FUNCTIONS are used to find the best relationship. In multiple regression, the dependent variable is considered to depend on more than a single independent variable.
Transport proteins that carry specific substances in the blood or across cell membranes.
A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.
The production of offspring by selective mating or HYBRIDIZATION, GENETIC in animals or plants.
A quality of cell membranes which permits the passage of solvents and solutes into and out of cells.

Induction of AT-specific DNA-interstrand crosslinks by bizelesin in genomic and simian virus 40 DNA. (1/6694)

Bizelesin is a bifunctional AT-specific DNA alkylating drug. Our study characterized the ability of bizelesin to induce interstrand crosslinks, a potential lethal lesion. In genomic DNA of BSC-1 cells, bizelesin formed from approx. 0.3 to 6.03+/-0.85 interstrand crosslinks per 106 base pairs, at 5-100 nM drug concentration, respectively, comparable to the number of total adducts previously determined in the same system (J.M. Woynarowski, M.M. McHugh, L.S. Gawron, T.A. Beerman, Biochemistry 34 (1995) 13042-13050). Bizelesin did not induce DNA-protein crosslinks or strand breaks. A model defined target, intracellular simian virus 40 (SV40) DNA, was employed to map at the nucleotide level sites of bizelesin adducts, including potential interstrand crosslinks. Preferential adduct formation was observed at AT tracts which are abundant in the SV40 matrix associated region and the origin of replication. Many sites, including each occurrence of 5'-T(A/T)4A-3', co-mapped on both DNA strands suggesting interstrand crosslinks, although monoadducts were also formed. Bizelesin adducts in naked SV40 DNA were found at similar sites. The localization of bizelesin-induced crosslinks in AT-rich tracts of replication-related regions is consistent with the potent anti-replicative properties of bizelesin. Given the apparent lack of other types of lesions in genomic DNA, interstrand crosslinks localized in AT-rich tracts, and to some extent perhaps also monoadducts, are likely to be lethal effects of bizelesin.  (+info)

The C-terminal region of hPrp8 interacts with the conserved GU dinucleotide at the 5' splice site. (2/6694)

A U5 snRNP protein, hPrp8, forms a UV-induced crosslink with the 5' splice site (5'SS) RNA within splicing complex B assembled in trans- as well as in cis-splicing reactions. Both yeast and human Prp8 interact with the 5'SS, branch site, polypyrimidine tract, and 3'SS during splicing. To begin to define functional domains in Prp8 we have mapped the site of the 5'SS crosslink within the hPrp8 protein. Immunoprecipitation analysis limited the site of crosslink to the C-terminal 5060-kDa segment of hPrp8. In addition, size comparison of the crosslink-containing peptides generated with different proteolytic reagents with the pattern of fragments predicted from the hPrp8 sequence allowed for mapping of the crosslink to a stretch of five amino acids in the C-terminal portion of hPrp8 (positions 1894-1898). The site of the 5'SS:hPrp8 crosslink falls within a segment spanning the previously defined polypyrimidine tract recognition domain in yPrp8, suggesting that an overlapping region of Prp8 may be involved both in the 5'SS and polypyrimidine tract recognition events. In the context of other known interactions of Prp8, these results suggest that this protein may participate in formation of the catalytic center of the spliceosome.  (+info)

Arginine methylation and binding of Hrp1p to the efficiency element for mRNA 3'-end formation. (3/6694)

Hrp1p is a heterogeneous ribonucleoprotein (hnRNP) from the yeast Saccharomyces cerevisiae that is involved in the cleavage and polyadenylation of the 3'-end of mRNAs and mRNA export. In addition, Hrplp is one of several RNA-binding proteins that are posttranslationally modified by methylation at arginine residues. By using functional recombinant Hrp1p, we have identified RNA sequences with specific high affinity binding sites. These sites correspond to the efficiency element for mRNA 3'-end formation, UAUAUA. To examine the effect of methylation on specific RNA binding, purified recombinant arginine methyltransferase (Hmt1p) was used to methylate Hrp1p. Methylated Hrp1p binds with the same affinity to UAUAUA-containing RNAs as unmethylated Hrpl p indicating that methylation does not affect specific RNA binding. However, RNA itself inhibits the methylation of Hrp1p and this inhibition is enhanced by RNAs that specifically bind Hrpl p. Taken together, these data support a model in which protein methylation occurs prior to protein-RNA binding in the nucleus.  (+info)

Photocrosslinking of 4-thio uracil-containing RNAs supports a side-by-side arrangement of domains 5 and 6 of a group II intron. (4/6694)

Previous studies suggested that domains 5 and 6 (D5 and D6) of group II introns act together in splicing and that the two helical structures probably do not interact by helix stacking. Here, we characterized the major Mg2+ ion- and salt-dependent, long-wave UV light-induced, intramolecular crosslinks formed in 4-thiouridine-containing D56 RNA from intron 5gamma (aI5gamma) of the COXI gene of yeast mtDNA. Four major crosslinks were mapped and found to result from covalent bonds between nucleotides separating D5 from D6 [called J(56)] and residues of D6 near and including the branch nucleotide. These findings are extended by results of similar experiments using 4-thioU containing D56 RNAs from a mutant allele of aI5gamma and from the group IIA intron, aI1. Trans-splicing experiments show that the crosslinked wild-type aI5gamma D56 RNAs are active for both splicing reactions, including some first-step branching. An RNA containing the 3-nt J(56) sequence and D6 of aI5gamma yields one main crosslink that is identical to the most minor of the crosslinks obtained with D56 RNA, but in this case in a cation-independent fashion. We conclude that the interaction between J(56) and D6 is influenced by charge repulsion between the D5 and D6 helix backbones and that high concentrations of cations allow the helices to approach closely under self-splicing conditions. The interaction between J(56) and D6 appears to be a significant factor establishing a side-by-side (i.e., not stacked) orientation of the helices of the two domains.  (+info)

Hairpin-shaped DNA duplexes with disulfide bonds in sugar-phosphate backbone as potential DNA reagents for crosslinking with proteins. (5/6694)

Convenient approaches were described to incorporate -OP(=O)O(-)-SS-O(-)(O=)PO- bridges in hairpin-shaped DNA duplexes instead of regular phosphodiester linkages: (i) H2O2- or 2,2'-dipyridyldisulfide-mediated coupling of 3'- and 5'-thiophosphorylated oligonucleotides on complementary template and (ii) more selective template-guided autoligation of a preactivated oligonucleotide derivative with an oligomer carrying a terminal thiophosphoryl group. Dithiothreitol was found to cleave completely modified internucleotide linkage releasing starting oligonucleotides. The presence of complementary template as an intrinsic element of the molecule protects the hairpin DNA analog from spontaneous exchange of disulfide-linked oligomer fragments and makes it a good candidate for auto-crosslinking with cysteine-containing proteins.  (+info)

Fibroblast growth factors 1 and 2 are distinct in oligomerization in the presence of heparin-like glycosaminoglycans. (6/6694)

Fibroblast growth factor (FGF) 1 and FGF-2 are prototypic members of the FGF family, which to date comprises at least 18 members. Surprisingly, even though FGF-1 and FGF-2 share more than 80% sequence similarity and an identical structural fold, these two growth factors are biologically very different. FGF-1 and FGF-2 differ in their ability to bind isoforms of the FGF receptor family as well as the heparin-like glycosaminoglycan (HLGAG) component of proteoglycans on the cell surface to initiate signaling in different cell types. Herein, we provide evidence for one mechanism by which these two proteins could differ biologically. Previously, it has been noted that FGF-1 and FGF-2 can oligomerize in the presence of HLGAGs. Therefore, we investigated whether FGF-1 and FGF-2 oligomerize by the same mechanism or by a different one. Through a combination of matrix-assisted laser desorption ionization mass spectrometry and chemical crosslinking, we show here that, under identical conditions, FGF-1 and FGF-2 differ in the degree and kind of oligomerization. Furthermore, an extensive analysis of FGF-1 and FGF-2 uncomplexed and HLGAG complexed crystal structures enables us to readily explain why FGF-2 forms sequential oligomers whereas FGF-1 forms only dimers. FGF-2, which possesses an interface capable of protein association, forms a translationally related oligomer, whereas FGF-1, which does not have this interface, forms only a symmetrically related dimer. Taken together, these data show that FGF-1 and FGF-2, despite their sequence homology, differ in their mechanism of oligomerization.  (+info)

The crayfish plasma clotting protein: a vitellogenin-related protein responsible for clot formation in crustacean blood. (7/6694)

Coagulation in crayfish blood is based on the transglutaminase-mediated crosslinking of a specific plasma clotting protein. Here we report the cloning of the subunit of this clotting protein from a crayfish hepatopancreas cDNA library. The ORF encodes a protein of 1,721 amino acids, including a signal peptide of 15 amino acids. Sequence analysis reveals that the clotting protein is homologous to vitellogenins, which are proteins found in vitellogenic females of egg-laying animals. The clotting protein and vitellogenins are all lipoproteins and share a limited sequence similarity to certain other lipoproteins (e.g., mammalian apolipoprotein B and microsomal triglyceride transfer protein) and contain a stretch with similarity to the D domain of mammalian von Willebrand factor. The crayfish clotting protein is present in both sexes, unlike the female-specific vitellogenins. Electron microscopy was used to visualize individual clotting protein molecules and to study the transglutaminase-mediated clotting reaction. In the presence of an endogenous transglutaminase, the purified clotting protein molecules rapidly assemble into long, flexible chains that occasionally branch.  (+info)

A region of the Yersinia pseudotuberculosis invasin protein enhances integrin-mediated uptake into mammalian cells and promotes self-association. (8/6694)

Invasin allows efficient entry into mammalian cells by Yersinia pseudotuberculosis. It has been shown that the C-terminal 192 amino acids of invasin are essential for binding of beta1 integrin receptors and subsequent uptake. By analyzing the internalization of latex beads coated with invasin derivatives, an additional domain of invasin was shown to be required for efficient bacterial internalization. A monomeric derivative encompassing the C-terminal 197 amino acids was inefficient at promoting entry of latex beads, whereas dimerization of this derivative by antibody significantly increased uptake. By using the DNA-binding domain of lambda repressor as a reporter for invasin self-interaction, we have demonstrated that a region of the invasin protein located N-terminal to the cell adhesion domain of invasin is able to self-associate. Chemical cross-linking studies of purified and surface-exposed invasin proteins, and the dominant-interfering effect of a non-functional invasin derivative are consistent with the presence of a self-association domain that is located within the region of invasin that enhances bacterial uptake. We conclude that interaction of homomultimeric invasin with multiple integrins establishes tight adherence and receptor clustering, thus providing a signal for internalization.  (+info)

TY - JOUR. T1 - Reversible DNA-Protein Cross-Linking at Epigenetic DNA Marks. AU - Ji, Shaofei. AU - Shao, Hongzhao. AU - Han, Qiyuan. AU - Seiler, Christopher L.. AU - Tretyakova, Natalia Y.. N1 - Funding Information: We thank Xun Ming and Peter Villalta (University of Minnesota) for their help with MS analysis and Robert Carlson (University of Minnesota) for preparing graphics for this manuscript. This research was supported by a grant from the NIEHS (ES023350). S.J. was partially supported by a Wayland E. Noland fellowship from the University of Minnesota. Publisher Copyright: © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. PY - 2017/11/6. Y1 - 2017/11/6. N2 - 5-Formylcytosine (5fC) is an endogenous DNA modification frequently found within regulatory elements of mammalian genes. Although 5fC is an oxidation product of 5-methylcytosine (5mC), the two epigenetic marks show distinct genome-wide distributions and protein affinities, suggesting that they perform different functions in ...
Homobifunctional crosslinkers possess identical reactive groups at opposite ends of the protein crosslinker s spacer arm, and can be reacted in a one-step chemical crosslinking reaction.
This invention is related to a thermoplastic crosslinked product obtainable by the crosslinking reaction of a composition comprising (A) a polymer having a silicon-containing group and (B) a tetravalent tin compound, said silicon-containing group having a hydrolyzable group bound to a silicon atom and capable of crosslinking through formation of a siloxane bond.
The CD4 receptor contributes to T-cell activation by coligating major histocompatibility complex class II on antigen presenting cells with the T-cell receptor (TCR)/CD3 complex, and triggering a cascade of signaling events including tyrosine phosphorylation of intracellular proteins. Paradoxically, CD3 cross-linking prior to TCR stimulation results in apoptotic cell death, as does injection of anti-CD4 antibodies in vivo of CD4 ligation by HIV glycoprotein (gp) 120. In this report we investigate the mechanism by which CD4 cross-linking induces cell death. We have found that CD4 cross-linking results in a small but rapid increase in levels of cell surface Fas, a member of the tumor necrosis factor receptor family implicated in apoptotic death and maintenance of immune homeostasis. Importantly, CD4 cross-linking triggered the ability of Fas to function as a death molecule. Subsequent to CD4 cross-linking, CD4+ splenocytes cultured overnight became sensitive to Fas-mediated death. Death was ...
In this work, intramolecular and intermolecular associations under the effect of shear flow of dilute aqueous alkaline solutions of dextan, hydroxyethylcellulose (HEC), and the hydrophobically modified analogue (HM-2-HEC) in the presence of a chemical cross-linking agent were characterized with the aid of viscometry and rheo-small angle light scattering (rheo-SALS) methods. The picture that emerges at short times during the cross-linking reaction at a constant shear rate is that HEC coils contract because of intramolecular cross-linking; whereas the HM-2-HEC species show an incipient association and the dextran molecules are unaffected due to their compact structures. At longer times, interchain cross-linking of the polymer promotes the growth of large flocs, which are disrupted by shear forces when they are sufficiently large. The delicate interplay between intramolecular and intermolecular association is governed by factors such as the magnitude of the shear rate, the cross-linking agent ...
DNA interstrand cross-links (ICLs) covalently link the Watson and Crick strands of DNA and are extremely cytotoxic. Widely used chemotherapeutics (e.g. nitrogen...
BACKGROUND: The biological effects of CD24 (FL-80) cross-linking on breast cancer cells have not yet been established. We examined the impact of CD24 cross-linking on human breast cancer cell line MCF-7. METHODS: MCF-7 and MDA-MB-231 cells were treated with anti-rabbit polyclonal IgG or anti-human CD24 rabbit polyclonal antibodies to induce cross-linking, and then growth was studied. Changes in cell characteristics such as cell cycle modulation, cell death, survival in three-dimensional cultures, adhesion, and migration ability were assayed after CD24 cross-linking in MCF-7. RESULTS: Expression of CD24 was analyzed by flow cytometry in MDA-MB-231 and MCF-7 cells where 2% and 66% expression frequencies were observed, respectively. CD24 cross-linking resulted in time-dependent proliferation reduction in MCF-7 cells, but no reduction in MDA-MB-231 cells. MCF-7 cell survival was reduced by 15% in three-dimensional culture after CD24 cross-linking. Increased MCF-7 cell apoptosis was observed after ...
Dried hemoactive materials comprise both a cross-linked biologically compatible polymer and a non-cross-linked biologically compatible polymer. The cross-linked polymer is selected to form a hydrogel when exposed to blood. The non-cross-linked polymer is chosen to solubilize relatively rapidly when exposed to blood. The non-cross-linked polymer serves as a binder for holding the materials in desired geometries, such as sheets, pellets, plugs, or the like. Usually, the cross-linked polymer will be present in a particulate or fragmented form. The materials are particularly suitable for hemostasis and drug delivery.
ChemInform Abstract: Incorporating Disulfide Cross-Links at the Terminus of Oligonucleotides via Solid-Phase Nucleic Acid Synthesis.|span||/span| | S. E. OSBORNE; A. D. ELLINGTON | download | BookSC. Download books for free. Find books
This chapter reviews modern trends in the development of cross-linking approaches and their implication for studies of rRNA folding in the ribosome and the organization of ribosomal functional centers. It illustrates the results emanating from the ribosomal cross-linking studies with data taken from the authors' work on the investigation of the environment of mRNA and 5S rRNA in the ribosome. The application of a combination of short-range and long-range crosslinking reagents has proved to be a very fruitful strategy for investigation of the topography of RNA and proteins in the ribosome. A section briefly describes the photo-reactive compounds used in most cross-linking experiments with ribosomes. Trifluoromethylaryldiazirines (TFMAD)-containing photoreagents can be used for scanning very fast conformational changes within the ribosome structure. The data reviewed in the chapter show that site-specific photo-cross-linking is a powerful tool for the identification of interacting elements of the
Choosing to participate in a study is an important personal decision. Talk with your doctor and family members or friends about deciding to join a study. To learn more about this study, you or your doctor may contact the study research staff using the contacts provided below. For general information, Learn About Clinical Studies. ...
The activation of G protein-coupled receptors (GPCR) mediates a variety of important biological signals. GPR110 belongs to the adhesion GPCR class with distinctively long N-terminus. Although GPR110 is an orphan receptor with unknown ligand, it has been identified as an oncogene implicated in lung and prostate cancers. In an effort to understand the molecular basis of GPR110 activation, we probed GPR110 conformation in living cells using chemical crosslinking and mass spectrometry. HEK cells overexpressing HA-GPR110 were incubated with disuccinimidyl suberate (DSS, a lysine-specific crosslinker). The DSS-modified HA-GPR110 was pulled-down and subjected to SDS-PAGE, tryptic digestion, and nanoLC-ESI-MS/MS. The MS-based approach identified 17 crosslinked lysine pairs in the N-terminal domain of GPR110. Among them, K29-K38, K187-K240, K240-K254, K398-442, K398-K438, K398-K427, K427-K442, K427-K438, and K151-K442, resulted from through-space crosslinking between two peptide segments, while K31-K32, ...
B. DNA-Protein Cross-Linking. To cross-link protein to the genomic DNA, 270 µl of formalin was added to 10 ml of DMEM containing acini to obtain a final solution of 1% formaldehyde. Cells were agitated for 10 minutes (Note 1) on a shaker table. Following the addition of 514 µl of 2.5 M glycine (125 mM final) for 5 minutes to quench the formaldehyde and terminate the crosslinking reaction, cells were centrifuged at 2000 rpm for 3 minutes at 40C. The pellet of acinar cells was washed twice with ice cold PBS.. C. Isolation of Nuclei from Isolated Acinar Cells. To isolate nuclei, acinar cells were incubated in Cytoplasmic Extract buffer (CE Buffer; 10 mM HEPES pH 7.4, 10 mM KCl, 1.5 mM MgCl2, 0.1% Triton-X100, 0.5 mM DTT and protease inhibitors [5 µg/mL Aprotinin, 5 µg/mL Leupeptin, 5 µg/mL Pepstatin, 75 µg/mL PMSF]) followed by 5 strokes of Potter-Elvehjem homogenizer. The cells were set on ice for 15 minutes. To pellet the nuclei, the samples were centrifuged at 4oC and 5000 g for 10 ...
Methods are provided for making bispecific antibodies and antibody conjugates comprising site-specifically cross-linking two or more antibodies, antibody fragments or Fc-fusion proteins. Also provided
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Cell lysis is an inevitable step in classical mass spectrometry-based strategies to analyse protein complexes. Complementary lysis conditions, in situ cross-linking strategies and proximal labelling techniques are currently used to reduce lysis effects on the protein complex. We have developed Virotrap, a viral particle sorting approach that obviates the need for cell homogenization and preserves the protein complexes during purification. By fusing a bait protein to the HIV-1 GAG protein, we show that interaction partners become trapped within virus-like particles (VLPs) that bud from mammalian cells. Using an efficient VLP enrichment protocol, Virotrap allows the detection of known binary interactions and MS-based identification of novel protein partners as well. In addition, we show the identification of stimulus-dependent interactions and demonstrate trapping of protein partners for small molecules. Virotrap constitutes an elegant complementary approach to the arsenal of methods to study protein
We have identified membrane components which are adjacent to type I and type II signal-anchor proteins during their insertion into the membrane of the ER. Using two different cross-linking approaches a 37-38-kD nonglycosylated protein, previously identified as P37 (High, S., D. Görlich, M. Wiedmann, T. A. Rapoport, and B. Dobberstein. 1991. J. Cell Biol. 113:35-44), was found adjacent to all the membrane inserted nascent chains used in this study. On the basis of immunoprecipitation, this ER protein was shown to be identical to the recently identified mammalian Sec61 protein. Thus, Sec61p is the principal cross-linking partner of both type I and type II signal-anchor proteins during their membrane insertion (this work), and of secretory proteins during their translocation (Görlich, D., S. Prehn, E. Hartmann, K.-U. Kalies, and T. A. Rapoport. 1992. Cell. 71:489-503). We propose that membrane proteins of both orientations, and secretory proteins employ the same ER translocation sites, and that ...
The 3′ major domain of Escherichia coli 16S rRNA, which occupies the head of the small ribosomal subunit, is involved in several functions of the ribosome. We have used a site-specific crosslinking procedure to gain further insights into the higher-order structure of this domain. Circularly permuted RNAs were used to introduce an azidophenacyl group at specific positions within the 3′ major domain. Crosslinks were generated in a high-ionic strength buffer that has been used for ribosome reconstitution studies and so enables the RNA to adopt a structure recognized by ribosomal proteins. The crosslinking sites were identified by primer extension and confirmed by assessing the mobility of the crosslinked RNA lariats in denaturing polyacrylamide gels. Eight crosslinks were characterized. Among them, one crosslink demonstrates that helix 28 is proximal to the top of helix 34, and two others show that the 1337 region, located in an internal loop at the junction of helices 29, 30, 41, and 42, is ...
SIA crosslinker is a non-cleavable, heterobifunctional protein crosslinker. SIA crosslinking reagent is the shortest amine and sulfhydryl (thiol) reactive protein crosslinker.
Using chemical cross-linkers to map contacts among amino acids, structural biologists predict that soluble tau is, in fact, a compact globule containing β-sheets poised to snap into a pathological formation.. ...
Using chemical cross-linkers to map contacts among amino acids, structural biologists predict that soluble tau is, in fact, a compact globule containing β-sheets poised to snap into a pathological formation.. ...
Hello, I am attempting to perform some experiments that inwolve cross-linking with Pierce photoreactive bifunctional cross-linker APDP. As I have no practice using that reagent, any advice would be most welcome. I plan to label one protein via sulphydryl end of APDP and then use the labelled protein in a reaction. After that the glycerol gradient will be run, and appropriate fractions will be flashed with light to activate the photoreactive group of APDP and cross-link that protein to whatever it has in proximity. I have read the papers provided by Pierce, but still have some questions, as follows: - Does APDP decompose thermally readily? Will it be possible to heat APDP-bound protein (that first one) for some 10 min, or will it destroy the photoreactive end (as I am affraid)? And how long can I keep APDP-bound protein sample before actual cross-linking with flashlight? - How photosensitive it is? Do I have to worry about ambient light when labelling the first protein by sulphydryl? - During the ...
Current hydrogels based on chondroitin sulfate (ChS) generally lack the necessary strength and precise mechanical tunability. Addressing these limitations, covalent cross-linking has evolved to produce hydrogels with desirable properties. However, such a methodology always precludes injection and self-healin
The present invention relates to a method of cross-linking a polymeric carbohydrate material with a second material by means of a soluble carbohydrate polymer and a crosslinking agent. The present invention furthermore relates to the resulting cross-linked material, to uses of the cross-linked material, as well as to a kit comprising the soluble carbohydrate polymer and the cross-linking agent. ...
Crosslinking is the process of chemically joining two or more molecules by a covalent bond. Crosslinking reagents contain two or more reactive ends that are capable of attaching to specific functional groups (primary amines, sulfhydryls, etc.) on proteins or other article describes some of the different crosslinkers and their use.
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Probing Akt-Inhibitor Interaction by Chemical Cross-Linking and Mass Spectrometry | Bill X. Huang; Hee-Yong Kim | download | BookSC. Download books for free. Find books
Various cross-linking methods have been developed for labeling of proteins (e.g. IgG) with fluorophores or enzyme (e.g. HRP). Cross-linking is the implementation of covalent bonds between functional groups of amino acids within a protein (intramolecular), or between interacting proteins (intermolecular) using chemical reagents.. Since the SERS-nanotags are indeed several thousand fold heavier than IgG, fluorophores or enzymes, particular attention must be paid to the chosen buffers and bioconjugation technique, which allows the linkage of bio- and nanomaterials [9]. Thus, for bioconjugation of SERS labels, gentle modification and adjustments are required. As it has been shown before, the stability and biofunctionality of SERS nanotags depends on the amount of crosslinker and the avoidance of centrifugation steps [9]. Figure 2C and D demonstrate two different preparations of anti p63 coupled SERS probes with and without the use of a reaction/separation chamber [9, 21]. The p63 IgG labeled SERS ...
Various cross-linking methods have been developed for labeling of proteins (e.g. IgG) with fluorophores or enzyme (e.g. HRP). Cross-linking is the implementation of covalent bonds between functional groups of amino acids within a protein (intramolecular), or between interacting proteins (intermolecular) using chemical reagents.. Since the SERS-nanotags are indeed several thousand fold heavier than IgG, fluorophores or enzymes, particular attention must be paid to the chosen buffers and bioconjugation technique, which allows the linkage of bio- and nanomaterials [9]. Thus, for bioconjugation of SERS labels, gentle modification and adjustments are required. As it has been shown before, the stability and biofunctionality of SERS nanotags depends on the amount of crosslinker and the avoidance of centrifugation steps [9]. Figure 2C and D demonstrate two different preparations of anti p63 coupled SERS probes with and without the use of a reaction/separation chamber [9, 21]. The p63 IgG labeled SERS ...
PRODUCTO ETX Cross-linking agent that improves attachment and wash fastness of resins. PRODUCTO R3 conc. Self-cross-linking agent of resin that improves wash and rubbing fastness of pigments.
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SF3B4 is one of four subunits of the splicing factor 3B. The protein cross-links to a region in the pre-mRNA immediately upstream of the branchpoint…
Tris-EDTA is used to dilute and store nucleic acid samples. Tris-EDTA-based solutions break protein cross-links and can therefore unmask antigens and epitopes in formalin-fixed and paraffin-embedded tissue sections. Treatment with TE Buffer enhances the staining intensity of antibodies in the immuno-histochemical detection of certain proteins.
There is provided herein a membrane or film comprising one or more aromatic ionomers covalently crosslinked through aryl-aryl (--Ar--Ar--), aryl-ether-aryl (--Ar--O--Ar--), aryl-sulfide-aryl (--Ar--S--Ar--), aryl-sulfone-aryl bonds, or any combination thereof, wherein said one or more aromatic ionomers further comprises at least one electron withdrawing group adapted to improve oxidant resistance of said membrane or film.
WESTBURY, NY--(Marketwired - September 11, 2015) - The crosslinking process utilizes short wave (254 nm) UV light to covalently bond nucleic acids to either nitrocellulose or nylon membranes. Spectronics Corporation manufactures advanced, versatile and accurate crosslinkers to avoid the loss of nucleic acids during the hybridization process, a consistent, irritating...
Bio-Synthesis offers peptide conjugation using various cross-linking chemistries. Carrier protein such as KLH, BSA, OVA can be conjugated to peptides.
Site-specific photocross-linking reveals that Sec61p and TRAM contact different regions of a membrane-inserted signal sequence. J Biol Chem. 1993 Dec 15; 268(35):26745-51 ...
Skin Factor 1 - 40000 - - AGE-breakerSkin Factor 1 is an AGE-breaker. AGE stands fornAdvanced-Glyvcation-Endproducts (AGE) and are crosslinked proteins and sugars. These
1XW7: Diabetes-associated mutations in human insulin: crystal structure and photo-cross-linking studies of a-chain variant insulin wakayama
The Spectrolinker XL-1000 UV crosslinker was made for applications including eliminating PCR contamination, cross-linking DNA an...
Molecular Cloning, also known as Maniatis, has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential.
When a low concentrations of crosslinker is added to resist (a film used to lay down the patterns of ever-shrinking lines and features on a chip, shown here at left), it is able to pattern smaller ...
TY - JOUR. T1 - Intramolecular crosslinking of hemoglobin A by sulfosuccinimidyl suberate. T2 - Application of crosslinked protein as a blood substitute. AU - Manjula, B. N.. AU - Acharya, A. S.. PY - 1994/11. Y1 - 1994/11. N2 - Sulfosuccinimidyl esters could be targeted to the residues of ββ cleft of hemoglobin A and the bis-sulfosuccinimidyl esters of aliphatic dicarboxylic acids could serve as the `affinity directed ββ crosslinkers of HbA. The reactivity of sulfosuccinimidyl esters of tartaric acid and sebacic acid with HbA was also investigated to establish the appropriate length of the alkyl chain optimal for the crosslinking reaction.. AB - Sulfosuccinimidyl esters could be targeted to the residues of ββ cleft of hemoglobin A and the bis-sulfosuccinimidyl esters of aliphatic dicarboxylic acids could serve as the `affinity directed ββ crosslinkers of HbA. The reactivity of sulfosuccinimidyl esters of tartaric acid and sebacic acid with HbA was also investigated to establish the ...
DNA interstrand cross-links (ICL) present a formidable challenge to the cellular DNA repair apparatus. For Escherichia coli, a pathway which combines nucleotide excision repair (NER) and homologous recombination repair (HRR) to eliminate ICL has been characterized in detail, both genetically and biochemically. Mechanisms of ICL repair in eukaryotes have proved more difficult to define, primarily as a result of the fact that several pathways appear compete for ICL repair intermediates, and also because these competing activities are regulated in the cell cycle. The budding yeast Saccharomyces cerevisiae has proven a powerful tool for dissecting ICL repair. Important roles for NER, HRR and postreplication/translesion synthesis pathways have all been identified. Here we review, with reference to similarities and differences in higher eukaryotes, what has been discovered to date concerning ICL repair in this simple eukaryote.
Chemical cross-linking was used to analyze the binding sites for the agonist bradykinin (BK) and the antagonists NPC17731 and HOE140 on the bovine B2 bradykinin receptor. [3H]BK and [3H]NPC17731 bound with high affinity to the same B2 receptor in bovine myometrial membranes as determined by the total number of specific binding sites and pharmacological specificity of the binding of these two radioligands. Cross-linking experiments were done using a series of bifunctional reagents reactive either primarily to amines (homobifunctional) or reactive to amines in one end and to sulfhydryls in the opposite end (heterobifunctional). All the heterobifunctional reagents plus the homobifunctional arylhalide 1,5-difluoro-2,4-dinitrobenzene were effective in cross-linking the [3H]BK N terminus specifically to a sulfhydryl in the receptor, and this cross-linking occurred at 5-100 μM reagent. In contrast, the homobifunctional N-hydroxysuccinimide ester reagents, at ≤1 mM, were only able to cross-link ...
For decades, formaldehyde has been routinely used to cross-link proteins in cells, tissue, and in some instances, even entire organisms. Due to its small size, formaldehyde can readily permeate cell walls and membranes, resulting in efficient cross-linking, i.e. the formation of covalent bonds between proteins, DNA, and other reactive molecules. Indeed, formaldehyde cross-linking is an instrumental component of many mainstream analytical/cell biology techniques including chromatin immunoprecipitation (ChIP) of protein-DNA complexes found in nuclei; immunohistological analysis of protein expression and localization within cells, tissues, and organs; and mass spectrometry (MS)-compatible silver-staining methodologies used to visualize low abundance proteins in polyacrylamide gels. However, despite its exquisite suitability for use in the analysis of protein environments within cells, formaldehyde has yet to be commonly employed in the directed analysis of protein-protein interactions and cellular ...
ABCA formed a novel intramolecular cross-linking adduct on human serum albumin in patients and in vitro via Michael addition, followed by nucleophilic adduction of the aldehyde with a neighbouring protein nucleophile. Adducts were detected on lysine, histidine, and cysteine residues in subdomain IB of human serum albumin. Only a cysteine adduct and a putative cross-linking adduct were detected on glutathione S-transferase Pi. Modelling the docking of ABCA with HLA B*57:01 confirmed that ABCA has a strong binding affinity when bound covalently to Ser116, a key residue with regards to recognition by ABC-specific CD8+ T cells. ...
215597-96-9,CCD No.:CCD00211814,Formula:C12 H10 N2 O14 S2 . 2 Na,MolWeight:516.323,Synonyms:BIS(SULFOSUCCINIMIDYL)SUCCINATE SODIUM SALT; Molecular Structure,Chemical Cloud Database
TY - JOUR. T1 - Efficient cross-linking to cytidine by functional nucleobases capable of in situ activation.. AU - Kawasaki, T.. AU - Nagatsugi, F.. AU - Usui, D.. AU - Maeda, M.. AU - Sasaki, S.. PY - 1999. Y1 - 1999. N2 - We have previously demonstrated that the ODNs with 2-amino-6-(2-phenylsulfoxyethyl)purine nucleoside derivative were capable of efficient interstrand cross-linking with cytidine selectively. In this new strategy, less reactive precursor was auto-activated within a duplex to generate 2-amino-6-vinylpurine derivative. However, it turned out that 2-amino-6-(2-phenylsulfinyl)-ethylpurine nucleoside was not applicable as the precursor for the synthesis of DNA oligomers with G-rich sequences. In this report, 2-amino-6-(2-methylsulfinylethyl)purine nucleoside has been proven to be more suitable as a precursor for DNA synthesis. In addition, the ODNs incorporating either 2-amino-6-(2-phenylsulfoxy ethyl)purine or 2-amino-6-vinylpurine showed high reactivity toward the cytidine at the ...
TY - JOUR. T1 - 4-vinyl-substituted pyrimidine nucleosides exhibit the efficient and selective formation of interstrand cross-links with RNA and duplex DNA. AU - Nishimoto, Atsushi. AU - Jitsuzaki, Daichi. AU - Onizuka, Kazumitsu. AU - Taniguchi, Yosuke. AU - Nagatsugi, Fumi. AU - Sasaki, Shigeki. PY - 2013/7. Y1 - 2013/7. N2 - The formation of interstrand cross-links in nucleic acids can have a strong impact on biological function of nucleic acids; therefore, many cross-linking agents have been developed for biological applications. Despite numerous studies, there remains a need for cross-linking agents that exhibit both efficiency and selectivity. In this study, a 4-vinyl-substituted analog of thymidine (T-vinyl derivative) was designed as a new cross-linking agent, in which the vinyl group is oriented towards the Watson-Crick face to react with the amino group of an adenine base. The interstrand cross-link formed rapidly and selectively with a uridine on the RNA substrate at the site opposite ...
TY - JOUR. T1 - Synthesis of peptide-oligonucleotide conjugates using a heterobifunctional crosslinker. AU - Williams, Berea A R. AU - Chaput, John C.. PY - 2010. Y1 - 2010. N2 - Peptide-oligonucleotide conjugates (POCs) are molecular chimeras composed of a nucleic acid moiety covalently attached to a polypeptide moiety. POCs have been used in numerous applications from therapeutics to nanotechnology, and most recently as combinatorial agents in the assembly of bivalent protein affinity reagents. This unit describes the synthesis and purification of POC molecules using the heterobifunctional crosslinking reagent succinimidyl-4-(N- maleimidomethyl)cyclohexane-1-carboxylate (SMCC), which enables amine-modified oligonucleotides to become covalently linked to cysteine-modified polypeptides. This solution-based protocol consists of a two-step synthesis followed by a single purification step.. AB - Peptide-oligonucleotide conjugates (POCs) are molecular chimeras composed of a nucleic acid moiety ...
Sigma-Aldrich offers bifunctional and trifunctional linkers and crosslinking reagents for various conjugation applications. There are numerous bioconjugation possibilities with our linkers and crosslinkers, offering potential for structural stability or assistance in protein-protein, protein-peptide and peptide/protein-small molecule interactions. Other applications include immobilization for assays or purification, as well as various peptide-nucleic acid and nucleic acid-nucleic acid conjugations, among many others. Our homo- and heterofunctional linkers contain diverse functional groups, such as primary amines, sulfhydryls, acids, alcohols and bromides. Many of our crosslinkers are functionalized with maleimide (sulfhydral reactive) and succinimidyl ester (NHS) or isothiocyanate (ITC) groups that react with amines. A selection of mono-protected (Boc, Fmoc, and Cbz) linkers is also available.
Reggie-1 and -2 proteins (flotillin-2 and -1 respectively) form their own type of non-caveolar membrane microdomains, which are involved in important cellular processes such as T-cell activation, phagocytosis and signalling mediated by the cellular prion protein and insulin; this is consistent with the notion that reggie microdomains promote protein assemblies and signalling. While it is generally known that membrane microdomains contain large multiprotein assemblies, the exact organization of reggie microdomains remains elusive. Using chemical cross-linking approaches, we have demonstrated that reggie complexes are composed of homo- and hetero-tetramers of reggie-1 and -2. Moreover, native reggie oligomers are indeed quite stable, since non-cross-linked tetramers are resistant to 8 M urea treatment. We also show that oligomerization requires the C-terminal but not the N-terminal halves of reggie-1 and -2. Using deletion constructs, we analysed the functional relevance of the three predicted ...
The DNA-binding inorganic compound cisplatin is one of the most successful anticancer drugs. The detailed mechanism by which cells recognize and process cisplatin−DNA damage is of great interest. Although the family of proteins that bind cisplatin 1,2- and 1,3-intrastrand cross-links has been identified, much less is known about cellular protein interactions with cisplatin interstrand cross-links (ICLs). In order to address this question, a photoreactive analogue of cisplatin, PtBP[subscript 6], was used to construct a DNA duplex containing a site-specific platinum ICL. This DNA probe was characterized and used in photo-cross-linking experiments to separate and identify nuclear proteins that bind to the ICL by peptide mass fingerprint analysis. Several such proteins were discovered, including PARP-1, hMutSβ, DNA ligase III, XRCC1, and PNK. The photo-cross-linking approach was independently validated by an electrophoretic mobility shift assay demonstrating hMutSβ binding to a cisplatin ICL. ...
Dear netter, I am trying to cross-link some of my mutated actin monomers within filament actin by using the zero-length cross-linker. Anybody has success with this cross-linker? Your protocol and advice will be highly appreciated. Bing ...
TY - JOUR. T1 - Sulfhydryl Site-Specific Cross-Linking and Labeling of Monoclonal Antibodies by a Fluorescent Equilibrium Transfer Alkylation Cross-Link Reagent. AU - del Rosario, Renato B.. AU - Wahl, Richard L.. AU - Brocchini, Stephen J.. AU - Lawton, Richard G.. AU - Smith, Richard H.. PY - 1990/1/1. Y1 - 1990/1/1. N2 - The site-specific intramolecular cross-linking of sulfhydryls of monoclonal antibodies via a new class of equilibrium transfer alkylation cross-link (ETAC) reagents is described. Following complete or partial reduction of interchain disulfides with dithiothreitol (DTT), two murine IgG2a monoclonal antibodies, 225.28S and 5G6.4, were reacted with a,a-bis[(p-tolylsulfonyl)methyl]-m-aminoacetophenone (ETAC 1a) and a fluorescent conjugated derivative, sulforhodamine B m-(α,α-bis(p-tolylsulfonylmethyl) acetyl)anilide derivative (ETAC 1b). Reducing SDS-polyacrylamide gel electrophoresis analysis of the products from lb indicated the formation of S-ETAC-S interchain heavy and ...
Thermo Scientific™ Pierce™ DSS Crosslinker 1g Thermo Scientific™ Pierce™ DSS Crosslinker NHS-ester Crosslinkers
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TY - CONF. T1 - Preparation of porous tubular scaffolds for vascular tissue engineering by photo-crosslinking of PTMC in a glass mold. AU - Guo, Zhengchao. AU - Grijpma, Dirk W.. AU - Poot, Andreas A.. PY - 2015/11/7. Y1 - 2015/11/7. KW - METIS-314723. M3 - Poster. T2 - International Conference on Biofabrication 2015. Y2 - 7 November 2015 through 9 November 2015. ER - ...
Enzymes are immobilized by dissolving in water a photo-crosslinking resin containing stilbazolium groups, vinyl alcohol units and vinyl acetate units, adding an enzyme to the resultant aqueous solution and exposing the enzyme-containing resin solution to light to induce a crosslinking reaction of the photo-crosslinking resin and produce a polymer containing the enzyme entrapped therein.
Shop a large selection of Crosslinking Reagents products and learn more about Thermo Scientific DSS (disuccinimidyl suberate), No-Weigh Format 50mg:Life 50mg.
It has been suggested that GAP-43 (growth-associated protein) binds to various proteins in growing neurons as part of its mechanism of action. To test this hypothesis in vivo, differentiated N1E-115 neuroblastoma cells were labeled with [35S]-amino acids and were treated with a cleavable crosslinking reagent. The cells were lysed in detergent and the lysates were centrifuged at 100,000 x g to isolate crosslinked complexes. Following cleavage of the crosslinks and analysis by two-dimensional gel electrophoresis, it was found that the crosslinker increased the level of various proteins, and particularly actin, in this pellet fraction. However, GAP-43 was not present, suggesting that GAP-43 was not extensively crosslinked to proteins of the cytoskeleton and membrane skeleton and did not sediment with them. GAP-43 also did not sediment with the membrane skeleton following nonionic detergent lysis. Calmodulin, but not actin or other proposed interaction partners, co-immunoprecipitated with GAP-43 from the
TY - JOUR. T1 - Purification and reconstitution of sterol transfer by native mouse ABCG5 and ABCG8. AU - Wang, Jin. AU - Zhang, Da Wei. AU - Lei, Ying. AU - Xu, Fang. AU - Cohen, Jonathan C.. AU - Hobbs, Helen H.. AU - Xie, Xiao Song. PY - 2008/5/6. Y1 - 2008/5/6. N2 - ABCG5 (G5) and ABCG8 (G8) are ATP-binding cassette half-transporters that limit intestinal uptake and promote biliary secretion of neutral sterols. Here, we describe the purification of endogenous G5G8 from mouse liver to near homogeneity. We incorporated the native proteins into membrane vesicles and reconstituted sterol transfer. Native gel electrophoresis, density-gradient ultracentrifugation, and chemical cross-linking studies indicated that the functional native complex is a heterodimer. No higher order oligomeric forms were observed at any stage in the catalytic cycle. Sterol transfer activity by purified native G5G8 was stable, stereospecific, and selective. We also report that G5 but not G8 is S-palmitoylated and that ...
Here we describe a protocol based mostly on structure-guided cysteine cross-linking and proteolysis-coupled gel evaluation to probe conformational modifications of a goal transporter in reside Escherichia coli cells. Although cross-linking approaches have been used to probe the proximity between transmembrane segments in membrane proteins in vivo, to our information this protocol is the primary for use to interrogate transporter dynamics in cells. The use of this protocol is perfect for proteins with recognized or modeled buildings to information the alternative of particular residues with cysteines and the choice of cross-linking brokers with varied spacer arm lengths. This protocol permits for discriminating simply cross-linked and uncross-linked species and doesnt require the customarily troublesome or unavailable reconstitution of transport exercise in an in vitro system. In addition, this protocol could possibly be used to probe the conformation of transporters in cells handled with ...
Summary. Background: Activated factor XIII (FXIIIa), a transglutaminase, introduces fibrin-fibrin and fibrin-inhibitor cross-links, resulting in more mechanically stable clots. The impact of cross-linking on resistance to fibrinolysis has proved challenging to evaluate quantitatively. Methods: We used a whole blood model thrombus system to characterize the role of cross-linking in resistance to fibrinolytic degradation. Model thrombi, which mimic arterial thrombi formed in vivo, were prepared with incorporated fluorescently labeled fibrinogen, in order to allow quantification of fibrinolysis as released fluorescence units per minute. Results: A site-specific inhibitor of transglutaminases, added to blood from normal donors, yielded model thrombi that lysed more easily, either spontaneously or by plasminogen activators. This was observed both in the cell/platelet-rich head and fibrin-rich tail. Model thrombi from an FXIII-deficient patient lysed more quickly than normal thrombi; replacement ...
Tissue transglutaminase (tTG) belongs to the family of transglutaminase enzymes that catalyze the posttranslational modification of proteins via Ca(2+)-dependent cross-linking reactions. The catalytic action of tTG results in the formation of an isopeptide bond that is of great physiological significance since it is highly resistant to proteolysis and denaturants. Although tTG-mediated cross-linking reactions have been implicated to play a role in diverse biological processes, the precise physiological function of the enzyme remains unclear. Recent data, however, suggest that the protein polymers resulting from tTG-catalyzed reactions may play a role in commitment of cells to undergo apoptosis. On the same token, tTG-mediated formation of insoluble protein aggregates may underlie the markers of numerous pathological conditions, such as the senile plaques in Alzheimers disease and the Lewy bodies in Parkinsons disease. In addition to catalyzing Ca(2+)-dependent cross-linking reactions, tTG can also
Tissue engineering typically requires a use of scaffolds when delivering tissue-specific cells to be engineered. Hydrogels are frequently used as scaffolds, because their composition, structure, and function resemble the natural tissue extracellular matrix. In this study, hyaluronate-alginate hybrid (HAH) was synthesized by conjugating alginate (ALG) with the hyaluronate (HA) backbone using various types of linkers. HAH hydrogel was prepared by physically cross-linking the HAH polymer in the presence of calcium ions without chemical cross-linkers. The mechanical stiffness of HAH hydrogel was significantly affected by changing the type of a linker between HA and ALG. The mechanical stiffness increased with increasing linker length, likely due to enhanced intermolecular reactions between HA and ALG. This enables controlling the mechanical properties of HAH hydrogels. The types of linkers used to synthesize HAHs also influenced the chondrogenic differentiation of ATDC5 cells cultured in HAH ...
2005 (English)In: 8th International Symposium of Polymers for Advanced Technologies, 13th-16th Sept. 2005, Budapest, Hungary, 2005Conference paper, Published paper (Other scientific) ...
These overt changes in the BM microenvironment most likely limit bloodstream formation in marrow as indicated by patchy hematopoiesis, and even more bloodstream cells are stated in various other organs, like the spleen.69 Lysyl oxidase (LOX) may stabilize collagen fibrils by covalent crosslinking70 and plays a part in solid tumor progression by matrix stiffening.53 Interestingly, megakaryocytes produced from principal myelofibrosis patients present upregulated LOX appearance, facilitating collagen crosslinking thereby.71 While direct ramifications of matrix technicians on the development of principal myelofibrosis remain unidentified, biomaterial strategies, such as for example minimal matrix types of scars68 and interpenetrating polymer systems,54 could be utilized to recapitulate key matrix compositions from the pathological marrow also to differ substrate stiffness independently. V.?BIOPHYSICAL Legislation OF Immune system CELLS WITH IMPLICATIONS IN Cancer tumor IMMUNOTHERAPY Constructed ...
aldehydes chemical defense food quality protein cross-linking protein modifications Weissflog, Jerrit; Adolph, Sven; Wiesemeier, Theresa; Pohnert, Georg
Polyamides comprising substantial amounts of Nylon-11 and/or Nylon-12 units are cross-linked by irradiation in the presence of an unsaturated cross-linking agent, preferably triallyl isocyanurate. The cross-linked products are particularly useful in the form of heat-recoverable shaped articles, e.g. heat-shrinkable tubing.
Various structural components of biological membranes are asymmetrically localized in the two surfaces of the membrane bilayer. This asymmetry is absolute for membrane (glyco) proteins, but only a partial asymmetry has been observed for membrane phospholipids. In the red cell membrane, choline-phospholipids are localized mainly in the outer monolayer whereas aminophospholipids are distributed almost exclusively in the inner monolayer. Several evidences are now available to suggest that this distribution of membrane phospholipids in red cells is directly or indirectly maintained by the membrane-associated cytoskeleton (membrane skeleton). This belief is well supported by the previous as well as recent studies carried out in the authors laboratory. Previously, it has been shown that lipid-lipid interactions play no major role in maintaining the transmembrane phospholipid asymmetry in erythrocytes, and that the asymmetry is lost upon covalent crosslinking of the major membrane skeletal protein, ...
In order for transplanted cells to survive in the body, they need to be gently implanted, as well as provide a suitable environment allowing them to attach, survive, grow and function. Because different cell types prefer different types of environments, the delivery matrix can be modified. The HyStem® hydrogels permit gentle implantation by supporting cells when they are injected through a needle. HyStem® hydrogels are highly customizable, allowing for a variation of gel stiffness, viscosity and gelation time. They are also biocompatible. BioTime has designed these hydrogels to dissolve when needed to gently release cells from the matrix.. In addition to cells, proteins and/or other biological factors it can also be used as a time-release depot for drugs or biological molecules.. The technology underlying BioTimes HyStem® hydrogels was developed at the University of Utah and is based on a unique chemical cross-linking strategy. Building upon this platform, BioTime has developed the HyStem ...
Microcapsules are formed in the absence of coacervation by providing an oil-in-water emulsion containing a polymeric, emulsifying agent having cross-linkable groups or complexing sites and admixing with the emulsion a cross-linking agent or a complexing agent which forms an impermeable coating around the dispersed oil droplets. The emulsifying agent may be non-proteinaceous or the protein, gelatin. Impermeable capsule walls are formed solely by the addition of the cross-linking or complexing agent and extraneous hardening agents are obviated. Moreover, the emulsifying agent may be a preformed, polymeric, cross-linking agent which eliminates the need for any separate cross-linking agent.
A number of clinically important antitumor agents such as cisplatin, cyclophosphamide (a nitrogen mustard) or carmustine (BCNU, a chloro ethyl nitroso urea) for...
556 The pyrrolo[2,1-c][1,4]benzodiazepines (PBDs) are naturally occurring antitumor antibiotics isolated from various Streptomyces species. They bind covalently in the minor groove of DNA at purine-guanine-purine motifs. SJG-136, a pyrrolobenzodiazepine dimer based on two PBD units joined through their C8-positions via a propyldioxy linker, is presently in Phase I evaluation in both the UK (through Cancer Research UK) and the USA (through the NCI) as a result of its striking in vitro and in vivo activity. As it contains two electrophilic centers, SJG-136 forms interstrand cross-links between guanines on opposite strands of DNA at Purine-GATC-Pyrimidine sequences. More recently we have explored the synthesis and biological activity of C2-aryl substituted PBD monomers that monoalkylate guanines in the minor groove of DNA rather than form cross-links. In this respect, their mechanism of action is similar to Ecteinascidin-743 (Yondelis, ET-743). We have found that these molecules possess encouraging ...
Mitoxantrone is an anticancer anthracenedione that can be activated by formaldehyde to generate covalent drug-DNA adducts. Despite their covalent natu
5-(hexyloxy)psoralen: forms a conjugate with oligonucleotide primers (at its omega-hexyl position) to promote site-specific photo-induced cross-linking at the end of the DNA fragment of interest before analysis by DGGE
Tim23p is imported via the TIM (translocase of inner membrane)22 pathway for mitochondrial inner membrane proteins. In contrast to precursors with an NH 2 -terminal targeting presequence that are imported in a linear NH 2 -terminal manner, we show that Tim23p crosses the outer membrane as a loop before inserting into the inner membrane. The Tim8p-Tim13p complex facilitates translocation across the intermembrane space by binding to the membrane spanning domains as shown by Tim23p peptide scans with the purified Tim8p-Tim13p complex and crosslinking studies with Tim23p fusion constructs. The interaction between Tim23p and the Tim8p-Tim13p complex is not dependent on zinc, and the purified Tim8p-Tim13p complex does not coordinate zinc in the conserved twin CX 3 C motif. Instead, the cysteine residues seemingly form intramolecular disulfide linkages. Given that proteins of the mitochondrial carrier family also pass through the TOM (translocase of outer membrane) complex as a loop, our study suggests ...
Crosslinkable compositions contain a calcium carbonate-rich filler coated with a carboxylic acid of the formula R.sup.4--(OCR.sup.3.sub.2--C(.dbd.O)OH).sub.y. The compositions exhibit low modulus, good adhesion to substrates, and low skin formation time.
To understand outcomes, traditional data collection will only take you so far. IQVIA offers methods and expertise that take you to where you want to be. Explore our Innovative data collection techniques applied to a wide range of sources, cross-linking data and closing gaps. Use classical observational, low interventional clinical studies, or novel hybrid approaches such as mosaic and enriched studies. Designed to generate actionable insights and give you validated, representative, and fast results. Discover our track record in EMA-accepted Drug Utilization Studies (DUS) in multiple therapy areas and geographies and our experience in working with the FDA, ENCePP, EUnetHTA , MHRA.. ...
Molecular Cloning, also known as Maniatis, has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential.
Cross-links are bonds that link one polymer chain to another. These bonds can either be covalent or ionic in nature. In polymer science, the use of cross-links to promote a difference in a polymers physical properties is referred to as cross-linking. When polymer chains are linked together by cross-links, they lose some of their ability to move as individual polymer chains. For example, a liquid polymer, which possesses freely flowing chains can be turned into a solid or gel by cross-linking the chains together. Cross-linking inhibits close packing of the polymer chains, preventing the formation of crystalline regions. The restricted molecular mobility of a cross-linked structure limits the extension of the polymer material under loading. This means that when a polymer is stretched, the cross-links prevent the individual chains from sliding past each other. In the process, the chains may straighten out, but once the stress is removed they return to their original position and the object ...
A highly stable lipase from Geobacillus thermocatenolatus (BTL2) and the enhanced green fluorescent protein from Aquorea victoria (EGFP) were recombinantly produced N-terminally tagged to the lectin domain of the hemolytic pore-forming toxin LSLa from the mushroom Laetiporus sulphureus. Such a domain (LSL 150), recently described as a novel fusion tag, is based on a β-trefoil scaffold with two operative binding sites for galactose or galactose-containing derivatives. The fusion proteins herein analyzed have enabled us to characterize the binding mode of LSL 150 to polymeric and solid substrates such as agarose beads. The lectin-fusion proteins are able to be quantitatively bound to both cross-linked and non-cross-linked agarose matrixes in a very rapid manner, resulting in a surprisingly dynamic protein distribution inside the porous beads that evolves from heterogeneous to homogeneous along the postimmobilization time. Such dynamic distribution can be related to the reversible nature of the ...
Cross-linking reagentsEdit. Cross-linking agents for proteins include:[citation needed] ... an injury-induced aggregation event that disulfide crosslinks proteins and facilitates their removal by plasmin". Cell Reports ...
Cross-linking reagentsEdit. Cross-linking agents for proteins include:[citation needed] ... "Nucleocytoplasmic coagulation: an injury-induced aggregation event that disulfide crosslinks proteins and facilitates their ...
Peters, K.; Richards, F. M. (1977). "Chemical Cross-Linking: Reagents and Problems in Studies of Membrane Structure". Annual ... Richards FM, Knowles JR (1968). "Glutaraldehyde as a protein cross-linking reagent". Journal of Molecular Biology. 37 (1): 231- ... In the 1970s, with a succession of students and postdocs, the lab developed a series of chemical, photochemical, and cross-link ... doi:10.1016/S0021-9258(19)67984-6. Quiocho FA, Richards FM (1964). "Intermolecular cross linking of a protein in the ...
Selective crosslinking of proteins to CDAP-activated polysaccharides. Vaccine. 18:1273, 2000. Shafer, D. E, J. K. Inman, and A ... John Inman, Lees learned bioconjugation, the linking together of biologically relevant molecules to increase their utility. In ... Activation of soluble polysaccharides with CDAP for use in protein-polysaccharide conjugate vaccines and immunological reagents ... Versatile and efficient synthesis of protein-polysaccharide conjugate vaccines using aminooxy reagents and oxime chemistry. ...
Futaki S. and Kitagawa K. (1994). "Peptide-Unit Assembling Using Disulfide Cross-Linking - a New Approach for Construction of ... doi:10.1016/0040-4039(94)88040-9. "Special Reagents for Thiol Groups". Aldrichimica Acta. 4 (3): 33-46. Thalmann A., Oertle K. ...
The dicarboxylic acids, especially the small and linear ones, can be used as crosslinking reagents. Dicarboxylic acids where ... The PubChem links gives access to more information on the compounds, including other names, ids, toxicity and safety. Adipic ...
... while sulfenamides are used extensively in the vulcanization process to assist cross-linking. Thiocyanates, R−S−CN, are related ... They are typically prepared by the reaction of amides with Lawesson's reagent. Isothiocyanates, with formula R−N=C=S, are found ... Disulfides R−S−S−R with a covalent sulfur to sulfur bond are important for crosslinking: in biochemistry for the folding and ... Sulfonic acids like trifluoromethanesulfonic acid is a frequently used reagent in organic chemistry. Sulfinic acids have ...
... is used as a crosslinker for the synthesis of cleavable photo-cross-linking reagent. Mono-BOC-cystamine is ... Peter E Nielsen, John B Hansen and Ole Buchard (1984). "Photochemical cross-linking of protein and DNA in chromatin". Biochem J ... Biotin-Peg2-Amine Biotin-Peg2-Maleimide Biotin-Peg3-Amine Biotin-Peg4-NHS Mono BOC Cystamine[permanent dead link], Anachem ...
"Methyl 4-mercaptobutyrimidate as a cleavable cross-linking reagent and its application to the Escherichia coli 30S ribosome". ... "Traut's Reagent Instructions" (PDF). Thermo Scientific. Retrieved 2015-07-08. CS1 maint: discouraged parameter (link). ... 2-Iminothiolane is a cyclic thioimidate compound also known as Traut's reagent. It is a thiolating reagent that reacts with ... Thus it allows for crosslinking or labeling of molecules such as proteins through use of disulfide or thioether conjugation. It ...
Sulfur forms cross-linking bridges between sections of polymer chains which affects the mechanical and electronic properties. ... The cure package consists of various reagents that modify the kinetics and chemistry of crosslinking. These include accelerants ... They act to boost the cure speed and increase cross-link density, but also shorten the induction time, which can lead to ... Ultimately, activators promote the efficient use of sulfur to give a high density of cross-links. Due to the low solubility of ...
Cross-linking the protein precursor of marine mussel adhesives: bulk measurements and reagents for curing. Langmuir 20 (9), ... The Fe(L-DOPA3) complex can itself account for much cross-linking and cohesion in mussel plaque, but in addition the iron ... 1999;80(1). Sever M.J.; Weisser, J.T.; Monahan, J.; Srinivasan, S.; Wilker, J.J. (2004) Metal-mediated cross-linking in the ... Proteins in the oothecal foam of the mantis are cross-linked covalently by small molecules related to L-DOPA via a tanning ...
... thiol modification and cross-linking reagents". Biochimica et Biophysica Acta (BBA) - Bioenergetics. 1458 (2-3): 443-56. doi: ... This link is tenuous, however, as the overall structure of flagellar motors is far more complex than that of the FO particle ... Cross RL, Taiz L (January 1990). "Gene duplication as a means for altering H+/ATP ratios during the evolution of FOF1 ATPases ... Cross RL, Müller V (October 2004). "The evolution of A-, F-, and V-type ATP synthases and ATPases: reversals in function and ...
... thiol modification and cross-linking reagents". Biochimica et Biophysica Acta (BBA) - Bioenergetics. 1458 (2-3): 443-56. doi: ... linked together by a central stalk and a peripheral stalk. As an A subunit, MT-ATP8 is contained within the non-catalytic, ...
Monahan, J.; Wilker, J.J. (2004) Cross-linking the protein precursor of marine mussel adhesives: bulk measurements and reagents ... The Fe(L-DOPA3) complex can itself account for much cross-linking and cohesion in mussel plaque,[10] but in addition the iron ... Proteins in the oothecal foam of the mantis are cross-linked covalently by small molecules related to L-DOPA via a tanning ... Sever M.J.; Weisser, J.T.; Monahan, J.; Srinivasan, S.; Wilker, J.J. (2004) Metal-mediated cross-linking in the generation of a ...
CS1 maint: discouraged parameter (link) (dead link 8 April 2018) Panuwet P, Wade EL, Nguyen JV, Montesano MA, Needham LL, Barr ... Testing for cyanuric acid concentration is commonly done with a turbidometric test, which uses a reagent, melamine, to ... Because of its trifunctionality, CYA is a precursor to crosslinking agents, especially for polyurethane resins and ... CS1 maint: discouraged parameter (link) CS1 maint: archived copy as title (link) M. Budnowski, Angew. Chem., 7, 827 (1968). " ...
It is also used as a cross-linking agent to cross-link chitosan membranes, and also improves the membrane's integrity. " ... Suberoyl chloride is used as a reagent to synthesize hydroxyferrocifen hybrid compounds that have antiproliferative activity ... CS1 maint: discouraged parameter (link) MSDS Safety data (PDF) v t e. ...
Cross-linking is the chemical process to produce textiles with wrinkle resistance.[4] Wrinkle resistance is achieved by the ... N-methylol and DMDHEU are reagents commonly used for the treatments because of their relatively low costs; however, they ... crosslinking of cellulose chains to stop the molecules from moving when in contact with water or other environmental stress.[5] ...
... the experimenter may apply SDS-PAGE after first treating the intact complex with chemical cross-link reagents. Proteins are ...
... is used as a solubilizer and cross-linking agent in polymer chemistry. Glyoxal is a valuable building block in organic ... A convenient form of the reagent for use in the laboratory is its bis(hemiacetal) with ethylene glycol, 1,4-dioxane-2,3-diol. ...
Photolithography can also be used to cross-link cell-seeded photo-polymerizable ECM for three-dimensional studies. Using ECM ... This approach uses less energy and has high throughput, but has large reagent consumption and gas bubbles can form inside the ... Other drawbacks of conventional PCR is the high consumption of expensive reagents, preference for amplifying short fragments, ... Digital PCR eliminates sample/reagent surface adsorption and contamination by carrying out PCR in microdroplets or ...
He realized that while the disulfide bonds, which stabilize proteins by cross-linking, were broken, the structures containing ... Scientist David R. Goddard, in the early 1930s, identified TGA as a useful reagent for reducing the disulfide bonds in proteins ...
... much less toxic than glutaraldehyde and many other commonly used synthetic cross-linking reagents. Furthermore, genipin can be ... Genipin is an excellent natural cross-linker for proteins, collagen, gelatin, and chitosan cross-linking. It has a low acute ... link) Brenda Vaandering, Genipin, retrieved 22 December 2019 Zhang, CY; Parton, LE; Ye, CP; Krauss, S; Shen, R; Lin, CT; Porco ...
External links[edit]. *Instant insight into cell penetrating peptides from the Royal Society of Chemistry ... However, the ability of cross caca membrane is not unidirectional; arginine-based CPPs are able to enter-exit the cell membrane ... This approach has greatly simplified the formulation of reagents.[58] As contrast agents transporters[edit]. ... The evaluation of cytosolic delivery of CPP linked proteins has been found to be prone to artifacts[52] and therefore requires ...
General Cross Linking". Journal of Chemical Physics. 12,4, 125 Sahini, M.; Sahimi, M. (2003-07-13). Applications Of Percolation ... the Flory-Stockmayer Theory can be used to predict whether gelation is possible through analyzing the limiting reagent of the ... Flory-Stockmayer theory is a theory governing the cross-linking and gelation of step-growth polymers. The Flory-Stockmayer ... In other words, polymer chains are cross-linked with other polymer chains to form an infinitely large molecule, interspersed ...
Witkowski A, Joshi AK, Rangan VS, Falick AM, Witkowska HE, Smith S (April 1999). "Dibromopropanone cross-linking of the ... is largely based on the observations that the bifunctional reagent 1,3-dibromopropanone (DBP) is able to crosslink the active ... and a reinvestigation of the DBP crosslinking experiments revealed that the KS active site Cys161 thiol could be crosslinked to ... External links[edit]. *Fatty+Acid+Synthase at the US National Library of Medicine Medical Subject Headings (MeSH) ...
Synthesis of extended equilibrium transfer alkylating cross-link reagents and their use in the formation of macrocyclesStephen ... applicable for reactions betwixt allylic compounds and reagents like SOCl2. The synthetic utility can be extended to ...
... is a cross-linked polystyrene resin that carries a chloromethyl functional group. Merrifield resin is named ... These beads are allowed to swell in suitable solvents (ethyl acetate, DMF, DMSO), which then allows the reagents to substitute ...
Seven residues and longer are sufficient for cross-linking, leading to precipitation of the glycans with the Yariv ... 3-linked Gal units to make aggregates with the Yariv reagent. After translation, the AGP protein backbones are highly decorated ... The structure of the AG glycans consists of a backbone of β-1,3 linked galactose (Gal), with sidechains of β-1,6 linked Gal and ... The AG found in AGPs is of type II (type II AGs) - that is, a galactan backbone of (1-3)-linked β-D-galactopyranose (Galp) ...
... cross-linking reagents MeSH D27.720.470.410.360 - intercalating agents MeSH D27.720.470.410.505 - luminescent agents MeSH ... reagent strips MeSH D27.720.470.410.690 - reducing agents MeSH D27.720.470.410.700 - sulfhydryl reagents MeSH D27.720.470.410. ... indicators and reagents MeSH D27.720.470.410.080 - affinity labels MeSH D27.720.470.410.080.600 - photoaffinity labels MeSH ... D27.720.470.410.505.500 - fluorescent dyes MeSH D27.720.470.410.650 - radiopharmaceuticals MeSH D27.720.470.410.680 - reagent ...
... synthesis and a component in many car airbag systems Psoralen combined with ultraviolet radiation causes DNA cross-linking and ... Sodium azide, an azide salt that is a common reagent in organic ... Sex-Linked Recessive Lethal Test - Males from a strain with ... These ROS may result in the production of many base adducts, as well as DNA strand breaks and crosslinks. Deaminating agents, ... The first mutagens to be identified were carcinogens, substances that were shown to be linked to cancer. Tumors were described ...
A notable reagent is Schlosser's base, a mixture of n-butyllithium and potassium tert-butoxide. This reagent reacts with ... permanent dead link] *^ Sevov, S.C. "Zintl Phases", pp. 113-132 in Intermetallic Compounds, Principles and Practice: Progress, ... and cross-section towards neutron absorption.[10]:74 ... CS1 maint: BOT: original-url status unknown (link). *^ a b ... Buszek, Keith R. (2001) "Sodium Amalgam" in Encyclopedia of Reagents for Organic Synthesis, Wiley. doi:10.1002/047084289X.rs040 ...
The cross-coupling is carried out at room temperature with a base, typically an amine, such as diethylamine, that also acts as ... This results in the increased reactivity of the reagents and the ability of the reaction to be carried out at room temperature ... J., 13: 666-676, doi:10.1002/chem.200600574 CS1 maint: Multiple names: authors list (link) Vechorkin, O.; Barmaz, D.; Proust, V ... The Sonogashira reaction is a cross-coupling reaction used in organic synthesis to form carbon-carbon bonds. It employs a ...
CS1 maint: multiple names: authors list (link). *^ "New Drugs". Can Med Assoc J. 80 (12): 997-998. 1959. PMC 1831125. PMID ... This is because fluorescein is xylem-mobile and unable to cross plasma membranes, making it particularly useful in tracking ... In oligonucleotide synthesis, several phosphoramidite reagents containing protected fluorescein, e.g. 6-FAM phosphoramidite 2,[ ... External links[edit]. Wikimedia Commons has media related to Fluorescein.. *Absorption and Emission Spectra of Fluorescein in ...
CS1 maint: multiple names: authors list (link). *^ Herbst, Eric (May 12, 2005). "Chemistry of Star-Forming Regions". Journal of ... Solutions of substances in reagent bottles, including ammonium hydroxide and nitric acid, illuminated in different colors ... There are also several main cross-disciplinary and more specialized fields of chemistry.[76] ... It is often seen as linked to the quest to turn lead or another common starting material into gold,[5] though in ancient times ...
External links. Wikimedia Commons has media related to Minerals.. The Wikibook Historical Geology has a page on the topic of: ... When minerals react, the products will sometimes assume the shape of the reagent; the product mineral is termed a pseudomorph ... Penetration twins consist of two single crystals that have grown into each other; examples of this twinning include cross- ... CS1 maint: uses authors parameter (link). *^ Grotzinger, J.P.; et al. (January 24, 2014). "A Habitable Fluvio-Lacustrine ...
Newnham, R.E.; Cross, L. Eric (November 2005). "Ferroelectricity: The Foundation of a Field from Form to Function". MRS ... It is a common precipitant in protein crystallography and is also an ingredient in the Biuret reagent which is used to measure ... It is an ingredient of Fehling's solution (reagent for reducing sugars). It is used in electroplating, in electronics and ... Fieser, L. F.; Fieser, M., Reagents for Organic Synthesis; Vol.1; Wiley: New York; 1967, p. 983 ...
Two-Potato Clock - Science Kit and Boreal Laboratories[permanent dead link]. Accessed 10 April 2007. ... For example, disposable batteries often use a zinc "can" both as a reactant and as the container to hold the other reagents. If ... a crossed-out wheeled bin). This must cover at least 3% of the surface of prismatic batteries and 1.5% of the surface of ... In this example the two half-cells are linked by a salt bridge that permits the transfer of ions. ...
... or by cross-coupling with beta-amino organozinc reagents, or reacting a brominated arene with beta-aminoethyl organolithium ... The findings may be linked to the antidepressant effects of exercise.. *^ a b c d Berry MD (2007). "The potential of trace ... reagents, or by Suzuki cross-coupling.[24]. Detection in body fluidsEdit. This section needs expansion with: [5]. You can help ... CS1 maint: Multiple names: authors list (link) *^ Lynnes T, Horne SM, Prüß BM (2014). "ß-Phenylethylamine as a novel nutrient ...
Some ytterbium halides are used as reagents in organic synthesis. For example, ytterbium(III) chloride (YbCl3) is a Lewis acid ... The kinetic of excitations in ytterbium-doped materials is simple and can be described within the concept of effective cross- ... External linksEdit. Wikimedia Commons has media related to Ytterbium.. Look up ytterbium in Wiktionary, the free dictionary. ...
External links[edit]. Look up reproducibility in Wiktionary, the free dictionary.. *Artifact Evaluation for computer systems' ... inappropriate use of statistical tests and use of reagents that were not appropriately validated.[26] John P. A. Ioannidis ... Cross-sectional study vs. Longitudinal study, Ecological study. *Cohort study *Retrospective. *Prospective ... CS1 maint: Extra text (link). *^ a b Subcommittee E11.20 on Test Method Evaluation and Quality Control (2014), Standard ...
The links are intended only as a guide and explanation. For a full account of the work done by each Nobel Laureate, please see ... "for their development of the use of boron- and phosphorus-containing compounds, respectively, into important reagents in ... "for palladium-catalyzed cross couplings in organic synthesis"[110] ஐ-இச்சி நெகிழ்சி United States ... The links in this column are to articles (or sections of articles) on the history and areas of chemistry for which the awards ...
Folin-Ciocalteu reagent(英语:Folin-Ciocalteu reagent). *Oxygen radical absorbance capacity(英语:Oxygen radical absorbance capacity) ... placebo-controlled cross-over study. Br J Nutr. 2009, 102 (7): 1065-1074. PMID 19402938. doi:10.1017/S0007114509359127.. 引文格式1维 ... 引文格式1维护:显式使用等标签 (link) ... 引文格式1维护:显式使用等标签 (link
DNA testing has evolved tremendously in the last few decades to ultimately link a DNA print to what is under investigation. The ... thus reducing the reagent usage as well as cost.[114] In some instances researchers have shown that they can increase the ... with each base generating various time specific signals corresponding to the sequence of bases as they cross the pore which are ... According to the model, DNA is composed of two strands of nucleotides coiled around each other, linked together by hydrogen ...
Two DNA bases that are cross-linked by a nitrogen mustard. Different nitrogen mustards will have different chemical groups (R ... A Departure from Preformed Organometallic Reagents. Topics in Current Chemistry. 279. Springer Science & Business Media. pp. 25 ... CS1 maint: Multiple names: authors list (link). *^ a b Gray OM, McDonnell GV, Forbes RB (Aug 2006). "A systematic review of ... CS1 maint: Multiple names: authors list (link). *^ a b Gray O, McDonnell GV, Forbes RB (2004). Gray, Orla, ed. "Methotrexate ...
CS1 maint: Uses authors parameter (link). *^ G. S. Hiers and F. D. Hager (1941). "Anisole". Organic Syntheses.. ; Collective ... Many cross coupling reactions proceed via oxidative addition as well. Electrophilic alkylating agentsEdit. Electrophilic ... H. Perst, D. G. Seapy (2008). "Triethyloxonium Tetrafluoroborate". Encyclopedia of Reagents for Organic Synthesis. doi:10.1002/ ... External linksEdit. *Macrogalleria page on polycarbonate production. *Alkylating+agents at the US National Library of Medicine ...
CS1 maint: BOT: original-url status unknown (link). *^ a b c d e f Ryan KJ, Ray CG, eds. (2004). Sherris Medical Microbiology ( ... Instrumentation can control sampling, reagent use, reaction times, signal detection, calculation of results, and data ... permanent dead link] *^ Vaisberg, Horneck G.; Eschweiler, U.; Reitz, G.; Wehner, J.; Willimek, R.; Strauch, K. (1995). " ... Each of the links must be present in a chronological order for an infection to develop. Understanding these steps helps health ...
Amorphous As2S3 does not possess a layered structure but is more highly cross-linked. Like other glasses, there is no medium or ... Certain organic reagents, used in organic solvents, permit the high-etch selectivity required to produce high-aspect ratio ... Both forms feature polymeric structures consisting of trigonal pyramidal As(III) centres linked by sulfide centres. The sulfide ...
External linksEdit. Wikimedia Commons has media related to RNA interference.. Wikiversity has learning resources about RNA ... Extensive efforts in computational biology have been directed toward the design of successful dsRNA reagents that maximize gene ... siRNAs that are automatically checked for possible cross-reactivity. ... Although the cleavage process has been proposed to be linked to translation, translation of the mRNA target is not essential ...
Activating reagents include phosphoryl chloride[86] and peptide coupling reagents.[77] Lysergic acid is made by alkaline ... CS1 maint: bot: original URL status unknown (link) *^ a b Krebs TS, Johansen PØ (July 2012). "Lysergic acid diethylamide (LSD) ... Tolerance to LSD builds up with consistent use[57] and cross-tolerance has been demonstrated between LSD, mescaline,[58] and ... LSD can be detected using an Ehrlich's reagent and a Hofmann's reagent. ...
CS1 maint: archived copy as title (link). *^ a b Margarete Seeger; Walter Otto; Wilhelm Flick; Friedrich Bickelhaupt; Otto S. ... At no point along its route does the line cross a permanent freshwater watercourse, so bore water had to be relied on. No ... "Highly Reactive Magnesium for the Preparation of Grignard Reagents: 1-Norbornane Acid", Organic Syntheses, Collected Volume 6, ...
... yet most problematic reagents with a tremendous number of factors that must be controlled in any experiment including cross ... External links[edit]. Wikimedia Commons has media related to Antibodies.. *Mike's Immunoglobulin Structure/Function Page at ... 1994). "Enzyme-linked immunosorbent assay (ELISA)". Methods Mol Biol. 32: 461-466. doi:10.1385/0-89603-268-X:461. ISBN 0-89603- ... Relatively weak binding also means it is possible for an antibody to cross-react with different antigens of different relative ...
External linksEdit. Wikiquote has quotations related to: Marcellin Berthelot. Wikimedia Commons has media related to Marcellin ... In 1863 he became a member of the Académie Nationale de Médecine; he was also awarded the Grand Cross of the Legion of Honour. ... Berthelot's reagent. NotesEdit. .mw-parser-output .reflist{font-size:90%;margin-bottom:0.5em;list-style-type:decimal}.mw-parser ...
CS1 maint: bot: original URL status unknown (link) *^ Cope, A. C.; Bach, R. D. (1973). "trans-Cyclooctene". Organic Syntheses. ... Silver carbonate is also used as a reagent in organic synthesis such as the Koenigs-Knorr reaction. In the Fétizon oxidation, ... is useful in nuclear reactors because of its high thermal neutron capture cross-section, good conduction of heat, mechanical ... CS1 maint: bot: original URL status unknown (link) *^ Meisler, Andy (18 December 2005). "A Tempest on a Tea Cart". Los Angeles ...
S/D treatment utilizes readily available and relatively inexpensive reagents, but these reagents must be removed from the ... It is predominantly used in the blood plasma industry, by over 50 organizations worldwide and by the American Red Cross [1]. ... but the level of virus removal is dependent on the column composition and the reagents that are used in the process. It is also ...
The chromium(III) stabilizes the leather by cross linking the collagen fibers.[70] Chromium tanned leather can contain between ... Both the chromate and dichromate anions are strong oxidizing reagents at low pH:[16] ... Potassium dichromate is a chemical reagent, used as a titrating agent.[84] ... crosses cell membranes and bioconcentrates up the food chain. In contrast, the toxicity of trivalent chromium is very low, ...
External links[edit]. Wikimedia Commons has media related to Sellafield.. Official information[edit]. *Nuclear Decommissioning ... In EARP the effectiveness of the process is enhanced by the addition of reagents to remove the remaining soluble radioactive ... HVDC Cross-Channel. *HVDC Moyle. *HVDC Norway-UK. *Isle of Man to England Interconnector ... Brief description, with link to very detailed article. *^ "The Nuclear Businesses". Archived from the original on 27 September ...
External links[edit]. Wikisource has the text of the 1905 New International Encyclopedia article "Laboratory".. ... the validation of publications and cross-cutting activities (number and type of seminars). ... laboratory glassware such as the beaker or reagent bottle. *Analytical devices as HPLC or spectrophotometers ... CS1 maint: location (link) *^ Michael L. Matson; Jeffrey P. Fitzgerald; Shirley Lin (October 1, 2007). "Creating Customized, ...
This strategy allows the otherwise challenging cross-aldol reaction between two aldehydes. In general, cross-aldol reactions ... External linksEdit. *Chem 206, 215 Lecture Notes (2003, 2006) by D. A. Evans, A. G. Myers, et al., Harvard University (pp. 345 ... One approach, demonstrated by Evans, is to silylate the aldol adduct.[58][59] A silicon reagent such as TMSCl is added in the ... Crossed-aldol reactant controlEdit. The problem of "control" in the aldol addition is best demonstrated by an example. Consider ...
... cross linking reagent · Cross Linking Reagents · disulfide · globular protein · milk protein · monomer · ovalbumin · thiol ... Cross-linked 2-nitroalkyl starches · Starch, cross-linking · Starch · Chemical reaction · Controlled study · Cross linking · ... Cross-Linking Reagents · Molecular Sequence Data · Nitro Compounds · Starch · Viscosity Granular starch was cross-linked with 1 ... cross linking reagent · arthritis · articular cartilage · collagen synthesis · cross linking · disease course · inflammation · ...
XLMOD is an ontology for cross-linking mass spectrometry reagents and GC-MS derivatization reagents.. ... mass spectrometry cross-linking and derivatization reagents Last uploaded: March 15, 2019 ...
Protein cross-linking agents and protein modification reagents optimized to help researchers achieve highly efficient reactions ... Protein Cross-Linking & Protein Modification * Protein Cross-Linkers * Homobifunctional Cross-Linkers * Heterobifunctional ... Protein Cross-Linking & Protein Modification. G-Biosciences offer a wide range of tools for protein cross-linking and protein ... Our high-quality reagents help researchers consistently achieve the protein cross-linking and protein modification results they ...
Note: You clicked on an external link, which has been disabled in order to keep your shopping session open. ... Chapter 5-Crosslinking and Photoactivatable Reagents * *Introduction to Crosslinking and Photoactivatable Reagents-Section 5.1 ... Amine-Amine Crosslinking The scientific literature contains numerous references to reagents that form crosslinks between amines ... Protein-Protein Crosslinking Kit. Our Protein-Protein Crosslinking Kit (P6305) provides all of the reagents and purification ...
Synthesis and Applications of a Two-Step Reagent Forming Cross-links between Ribonucleic Acid and Protein GERALD FINK; GERALD ... GERALD FINK, RICHARD BRIMACOMBE; Synthesis and Applications of a Two-Step Reagent Forming Cross-links between Ribonucleic Acid ...
Mesh term Cross-Linking Reagents. Browse to parent terms:. Indicators and Reagents. Description. Reagents with two reactive ...
Reagents with two reactive groups, usually at opposite ends of the molecule, that are capable of reacting with and thereby ... What is cross-linking reagents?. 3 years ago by Pozzter Q&A 0 replies, read ~324032 times ... Reagents with two reactive groups, usually at opposite ends of the molecule, that are capable of reacting with and thereby ...
Cross-Linking Reagents, Linkers, Ligands. HOAT (1-Hydroxy-7-azobenzotriazole) , 98%, Trifluoroacetic Acid (TFA) , 99.9%, HATU ... Cross-Linking Reagents, Linkers, Ligands Coupling & Cross-Linking Reagents, Linkers, Ligands. 1-(3-Dimethylaminopropyl)-3-Ethyl ...
Comparison of the peptide maps obtained from digested cross-linked ParR dimers in the presence and absence of a thiol reagent ... The strategy of chemical cross-linking combined with differential MALDI-MS peptide mapping (+ thiol reagent) enabled ... thiol reagent). The data revealed the presence of an intermolecular cross-link between the receptor regions of the glycoprotein ... Glycoprotein fusion constructs CD28-IgG and CD80-Fab were cross-linked in vitro by DTSSP, characterized by nonreducing SDS-PAGE ...
Bulk quantities of DSS crosslinking reagent are also available from ProteoChem. - DSS crosslinker is a homobifunctional protein ... crosslinker that is cell membrane permeable and can be used to crosslink intracellular proteins. - ... DSS crosslinking reagent can be used to crosslink intracellular proteins. If extracellular crosslinking is desired, then BS3, ... Protein Crosslinking Reagents » Homobifunctional Crosslinkers » Disuccinimidyl suberate (DSS) » DSS Crosslinking Reagent 1 gram ...
Thermo Scientific™ EZ-Link™ NHS-PEG4-Biotin No-Weigh™ Format is a pegylated, water-soluble reagent for simple and efficient ... Thermo Scientific™ EZ-Link™ Sulfo-NHS-Biotin No-Weigh™ Format is a short-chain, water-soluble biotinylation reagent for ... Thermo Scientific™ EZ-Link™ Sulfo-NHS-LC-Biotin No-Weigh™ Format is an intermediate-length, water-soluble biotinylation reagent ... Thermo Scientific™ Pierce™ Traut's Reagent. 2-Iminothiolane-HCl (CAS 4781-83-3); an amine-reactive thiolation reagent to ...
Cross-linking reagentsEdit. Cross-linking agents for proteins include:[citation needed] ... an injury-induced aggregation event that disulfide crosslinks proteins and facilitates their removal by plasmin". Cell Reports ...
Learn more about protein crosslinking and browse popular protein crosslinking products amongst many categories including other ... Crosslinking is the process of chemically joining two or more molecules by a covalent bond. Crosslinking reagents contain ... Note: You clicked on an external link, which has been disabled in order to keep your shopping session open. ... Crosslinking reagents based on maleimide and hydrazide reactive groups for conjugation and formation of covalent crosslinks ...
Cross-linking reagentsEdit. Cross-linking agents for proteins include:[citation needed] ... "Nucleocytoplasmic coagulation: an injury-induced aggregation event that disulfide crosslinks proteins and facilitates their ...
An intramolecular crosslink occurs when both protein linking groups of the crosslinking reagent cross-link to accessible amino ... subject crosslinking reagents may be used to cross-link two members of such complexes. The length of the crosslinking reagent ... The crosslinking reagents include a trifunctional scaffold that links to two protein linking groups and also links to an ... The subject kits contain at least the crosslinking reagent. The kit may also contain reagents for cross-linking, affinity ...
After 4 days at 37°C, cell viability was measured by addition of Cell Titer Glow (Promega) reagent followed by luminescence ... to compare ADCs of a DNA-cross-linking IGN with ADCs of a DNA-alkylating IGN that would be incapable of forming cross-links, we ... with the corresponding cross-linking ADC 2a, toward cocultured FRα+ and FRα− cells. B, Export of IGN catabolites 1a (cross- ... lysine-linked) or approximately 2 IGN molecules linked per antibody (cysteine-linked), as determined by UV-Vis spectroscopy ...
The guanine and cytosine content correlates with the degree of mitomycin-induced cross-linking. At high concentrations of the ...
Cross-Linking and Labeling of Monoclonal Antibodies by a Fluorescent Equilibrium Transfer Alkylation Cross-Link Reagent. In: ... Cross-Linking and Labeling of Monoclonal Antibodies by a Fluorescent Equilibrium Transfer Alkylation Cross-Link Reagent. / del ... Cross-Linking and Labeling of Monoclonal Antibodies by a Fluorescent Equilibrium Transfer Alkylation Cross-Link Reagent. ... Cross-Linking and Labeling of Monoclonal Antibodies by a Fluorescent Equilibrium Transfer Alkylation Cross-Link Reagent, ...
Reagents.. Human recombinant GSTP1-1 was from Sigma-Aldrich (St. Louis, MO) or from Alpha Diagnostics International Inc. (San ... Cross-linking and inactivation of GSTP1-1 by cyPG could be exploited as a novel strategy to overcome cancer cell resistance to ... It is noteworthy that 15d-PGJ2 elicited GSTP1-1 cross-linking in vitro, a process that could be mimicked by other dienone ... Thus, according to these results, Cys101 is the only residue essential for 15d-PGJ2-induced GSTP1-1 cross-linking. We showed ...
In general the protein/DNA complex is fixed with formaldehyde, a cross-linking agent that, because of its short spacer arm ( ... Cross-linking Chromatin Immunoprecipitation (ChIP) has become a popular method to detect the in vivo binding of proteins to DNA ... Materials & Reagents. Cell Lysis Buffer. 5mM PIPES pH 8. 85mM KCl. 0.5% NP40. 1mM PMSF. Protease inhibitor cocktail (Roche). ... a cross-linking agent that, because of its short spacer arm (about 2 A), generates reversible covalent links, mainly between ...
Cross-Linking Reagents. Secondary source ID. *PDB/2FRP. *PDB/2FS3. *PDB/2FSY ... Mutants that cannot crosslink or EI-IV particles that have been rendered incapable of forming the final crosslink remain in the ... 7 laevo lattice with each subunit covalently crosslinked to two neighbors. Well-characterized pH 4 expansion intermediates make ... to the Head II conformation that is secured at this extent only by the formation of the final class of covalent crosslinks. ...
Peptide cross-links formed using the homobifunctional-linker diethyl suberthioimidate (DEST) are shown to be ETD-cleavable. ... Soderblom, E.J., Goshe, M.B.: Collision-induced dissociative chemical cross-linking reagents and methodology: applications to ... Digestion of Cross-linked Proteins. Proteomics grade trypsin was added to DEST cross-linked protein solutions at a 1:20 (w/w) ... For cross-links matched from ETD spectra, a minimum of two mass pair peaks was required. For cross-links found in CID spectra, ...
Drugs and Reagents. Cell culture reagents were purchased from Quality Biological, Inc. (Gaithersburg, MD) except for fetal ... DNA-protein cross-links. ISC. interstrand cross-links. CPT. camptothecin. GI50. 50% growth inhibition. top1. topoisomerase I. ... Figure5 demonstrates that NSC 652287 also induced DNA-DNA cross-links. Such DNA-DNA cross-links retard the elution of DNA that ... The present study shows that NSC 652287 induces both DPC and DNA-DNA cross-links but no DNA strand breaks. These cross-links ...
Iron-catalyzed cross-coupling between 1-Bromoalkynes and Grignard-derived organocuprate reagents ... Iron-catalyzed cross-coupling between 1-Bromoalkynes and Grignard-derived organocuprate reagents. European Journal of Organic ... A series of alkynylarenes were successfully synthesised by starting from different 1-bromoalkynes and Grignard reagents. The ... An efficient iron-catalyzed cross-coupling reaction between alkynyl bromides and Grignard-derived organocuprates has been ...
Cross-Linking Reagents * DNA Restriction Enzymes * Electrophoresis, Polyacrylamide Gel * Encephalomyocarditis virus / genetics ... UV-irradiation on extracts supplemented with in vitro synthesized 32P-labelled transcripts followed by analysis of crosslinked ...
Cross-Linking Reagents / pharmacology* * Enzyme Activation / drug effects * Extracellular Signal-Regulated MAP Kinases / ... Here, we developed a novel two-step chemical crosslinking strategy to overcome these problems which resulted in a dramatic ... None of these but one protein was detectable without crosslinking procedures. The present method provides a sensitive and ... Effective identification of Akt interacting proteins by two-step chemical crosslinking, co-immunoprecipitation and mass ...
... are shown to be a valuable means of characterizing cross-linking sites. Cisplatin is different from current cross-linking ... 2011) Mass spectrometry evidence for cisplatin as a protein cross-linking reagent. Analytical Chemistry, Vol.83 (No.13). pp. ... Thus, it has the potential to function as a cross-linker. In this report, the cross-linking ability of cisplatin is ... Cisplatin is a potent anti-cancer drug, which functions by cross-linking adjacent DNA guanine residues. However within one day ...
cross-linking reagent A reagent with two reactive groups, usually at opposite ends of the molecule, that are capable of ... methoxsalen (CHEBI:18358) has role cross-linking reagent (CHEBI:50684) methoxsalen (CHEBI:18358) has role dermatologic drug ( ...
Collision-induced dissociative chemical cross-linking reagents and methodology: Applications to protein structural ... have developed a novel set of collision-induced dissociative cross-linking reagents and methodology for chemical cross-linking ... Following verification and subsequent optimization of cross-linked peptide complex dissociation, our reagents were applied to ... Collision-induced dissociative chemical cross-linking reagents and methodology: Applications to protein structural ...
... cross-linking reagents would contain two functional groups, which react with target components, thus linking them together. The ... Monofunctional and Zero -Length Cross -Linking Reagents. From a logical point of view, cross-linking reagents would contain two ... Chemistry of Protein and Nucleic Acid Cross-Linking and Conjugation. DOI link for Chemistry of Protein and Nucleic Acid Cross- ... Chemistry of Protein and Nucleic Acid Cross-Linking and Conjugation. DOI link for Chemistry of Protein and Nucleic Acid Cross- ...
  • The high reactivity of thiols ( Thiol-Reactive Probes-Chapter 2 ) and-with the exception of a few proteins such as β-galactosidase-their relative rarity in most biomolecules make thiol groups ideal targets for controlled chemical crosslinking. (
  • DSS crosslinking reagent can be used to crosslink intracellular proteins. (
  • HABA dye reagent and biotinylated protein standard for convenient colorimetric determination of biotinylation levels in antibodies and other proteins. (
  • Thermo Scientific™ EZ-Link™ Sulfo-NHS-LC-Biotin No-Weigh™ Format is an intermediate-length, water-soluble biotinylation reagent for labeling antibodies, proteins and other molecules that have primary amines. (
  • Biotinylate proteins or surfaces using EDC or other amine-reactive crosslinkers with this short (20.4A) biotinylation reagent containing a 2-unit polyethylene glycol spacer. (
  • Thermo Scientific™ EZ-Link™ NHS-PEG4-Biotin No-Weigh™ Format is a pegylated, water-soluble reagent for simple and efficient biotin labeling of antibodies, proteins and other primary amine-containing macromolecules. (
  • Diazirine analogs of leucine and methionine to express proteins in cell culture that will crosslink their protein interactors upon UV-light activation in vivo. (
  • Crosslinking reagents contain reactive ends to specific functional groups, such as primary amines and sulfhydryls, on proteins or other molecules. (
  • The availability of several chemical groups in proteins and peptides make them targets for conjugation and for study using crosslinking methods. (
  • Aryl azide, diazirine, and other photo-reactive (light-activated) chemical heterobifunctional crosslinking reagents to conjugate proteins, nucleic acids and other molecular structures involved in receptor-ligand interaction complexes via two-step activation. (
  • Check out our various types of in vivo crosslinking reagents to target cell surface or intracellular proteins and to accommodate different downstream analysis methods. (
  • The distance between the two protein linking groups can be selected to crosslink two proteins of a protein complex via accessible amino acid residues. (
  • These reagents and methods find use in a variety of applications in which crosslinking of proteins in desired. (
  • Cross-linking Chromatin Immunoprecipitation (ChIP) has become a popular method to detect the in vivo binding of proteins to DNA. (
  • In general the protein/DNA complex is fixed with formaldehyde, a cross-linking agent that, because of its short spacer arm (about 2 A), generates reversible covalent links, mainly between proteins and DNA. (
  • Covalent links are reversed and proteins are removed from DNA through phenol/chloroform extraction. (
  • To specifically address this problem, we thought of combining the use of formaldehyde with other cross-linking agents to improve the formation of covalent links between proteins and stabilize the association of proteins to promoters, including those that are not directly bound to DNA. (
  • This was done using UV-irradiation on extracts supplemented with in vitro synthesized 32P-labelled transcripts followed by analysis of crosslinked proteins by SDS-polyacrylamide gel electrophoresis. (
  • A reagent with two reactive groups, usually at opposite ends of the molecule, that are capable of reacting with and thereby forming bridges between macromolecules, principally side chains of amino acids in proteins, allowing the locations of naturally reactive areas within the proteins to be identified. (
  • However, there are some compounds that do not belong to either of these two classes, but are still able to effect cross-linking of two proteins or other macromolecules. (
  • On the other hand, zero-length reagents induce direct joining of two intrinsic chemical groups of proteins or other molecules without introduction of any extrinsic material. (
  • Other reagents that are not suitable for cross-linking proteins, but which have been used to couple macromolecules to solid supports, will be discussed in a later chapter. (
  • To address this problem, binding sites for the snoRNP proteins Nop1, Nop56, Nop58, and Rrp9 were mapped by UV cross-linking and analysis of cDNAs. (
  • The snoRNP proteins were also cross-linked to pre-rRNA fragments, with preferential association at known sites of box C/D snoRNA function. (
  • RNA immunoprecipitation uses formaldehyde to cross-link RNA to proteins ( 1 , 2 ). (
  • Caveats of this method are that formaldehyde also cross-links proteins to proteins and the immunoprecipitation step is performed under semidenaturing conditions, so a positive result does not demonstrate direct RNA-protein interaction, and the spatial resolution of the technique is low. (
  • This problem can be avoided by cross-linking proteins and RNA with UV light, and several techniques have been reported ( 3 ⇓ ⇓ ⇓ - 7 ). (
  • To analyze snoRNP and preribosome structures we established UV cross-linking methods and used them to map the binding sites for U3 snoRNP proteins on the snoRNA and the pre-rRNA. (
  • Chemical cross-linking of reactive groups in native proteins and protein complexes in combination with the identification of cross-linked sites by mass spectrometry has been in use for more than a decade. (
  • In the last decade, the application of protein cross-linking has expanded, first and foremost driven by developments in mass spectrometry as the method of choice for the high throughput identification of proteins and their modifications. (
  • Here, we present an overview of recent developments in methodology, instrumentation, and bioinformatics related to chemical cross-linking of proteins and the analysis of cross-linked peptides by mass spectrometry. (
  • To study the potential of the tyrosine click reaction to introduce poly(ethylene) glycol chains onto proteins (PEGylation), we demonstrate that this novel reagent provides for the selective PEGylation of chymotrypsinogen whereas traditional succinimide-based PEGylation targeting lysine residues provided a more diverse range of PEGylated products. (
  • Organized into five chapters, the book starts by reviewing the methods available for the immobilization of proteins, which includes enzymes adsorption, occlusion in cross-linked polymeric matrices, covalent binding to water-insoluble carriers, and intermolecular cross-linking. (
  • Crosslink sulfhydryls to sulfhydryls (cysteine residues) with this long, cleavable crosslinker composed of maleimide groups and 11-atom (13.3A) disulfide spacer arm. (
  • Cisplatin is a potent anti-cancer drug, which functions by cross-linking adjacent DNA guanine residues. (
  • However, unambiguous identification of the residues involved in a cross-link remains analytically challenging. (
  • Cross-linked residues identified by our CID-CXL-MS/MS method were in agreement with published crystal structures and previous cross-linking studies using conventional approaches. (
  • For this reason, intramolecular cross-linking can only occur between very closely juxtaposed residues in a macromolecular complex. (
  • 1 ) used chemical cross-linking of lysine residues in bovine basic fibroblast growth factor FGF-2 (heparin-binding growth factor 2) to provide distance constraints for the computational derivation of the fold of this small (17-kDa) protein. (
  • The major aim of the cross-linking reaction is the formation of a covalent bond between two spatially proximate residues within a single or between two polypeptide chains. (
  • Chemical cross-linking, followed by identification of the cross-linked residues, is a powerful approach to probe the topologies and interacting surfaces of protein assemblies. (
  • Mass spectrometric identification of the cross-linked residues provides valuable spatial restraints in defining protein interacting surfaces. (
  • The difficulty is largely due to the fact that most generic cross-linking reagents react with multiple residues and meanwhile undergo competing side reactions (with solvent, for example), inactivating one of the reactive groups. (
  • The lysine residues of Type 1 porcine collagen were directly cross-linked using l,4-Butanediol diglycidyl ether (BDDGE) under basic conditions at pH 11. (
  • and-like all of the thiolation reagents in this section-to introduce the highly reactive thiol group into peptides, onto cell surfaces or onto affinity matrices for subsequent reaction with fluorescent, enzyme-coupled or other thiol-reactive reagents ( Thiol-Reactive Probes-Chapter 2 ). (
  • Cleavable cross-linkers were introduced to reduce some of the above problems through the formation of diagnostic fragmentation patterns, such as reporter ions and peak doublets associated with constituent peptides [ 17 ]. (
  • Bis-arylhydrazone (BAH) cross-linked peptides cleave at the N-N hydrazone bond yielding one radical peptide and one even-electron peptide. (
  • DEB cross-linked peptides maintain the two protonation sites of amino moieties and yield diagnostic ions that reveal the intact unmodified constituent peptides [ 42 ]. (
  • In this report, the cross-linking ability of cisplatin is demonstrated by Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS) with the use of standard peptides, the 16.8 kDa protein calmodulin (CaM), but was unsuccessful for the 64 kDa protein hemoglobin. (
  • The high resolution and mass accuracy of FTICR MS along with the high degree of fragmentation of large peptides afforded by collisionally activated dissociation (CAD) and electron capture dissociation (ECD) are shown to be a valuable means of characterizing cross-linking sites. (
  • The success of XLMS relies on the ability to obtain high coverage of cross-linked peptides which are then used as restraints to guide modeling efforts. (
  • We next cross-link the yeast proteasome, identifying 3,893 unique cross-linked peptides in 3 mass spectrometry runs. (
  • Cross-linked peptides were separated by HPLC and analyzed on line by ESI-TOF and off line by MALDI-TOF mass spectrometry. (
  • In this work, we demonstrate a new bioinformatics approach using multiple program modules within the software package "Protein Prospector" that greatly facilitates the discovery of cross-linked peptides in chemical cross-linking studies. (
  • This means that the dominant reaction products are where only one end of the reagent reacts with a peptide, and these are referred to as dead-end modified peptides ( 5 ). (
  • However, mere incorporation of these moieties cannot distinguish the cross-linked peptides fromdead-end modified products ( 16 ). (
  • The high complexity of these samples requires tandem mass spectrometric fragmentation data for confident identification of the cross-linked peptides. (
  • Thus far, most of the reported software either rely on prescreening for cross-linker-containing peptides by their distinct isotope pattern ( 17 - 19 ) or require manual entry of peak lists for each predetermined cross-linked peptide candidate ( 5 ). (
  • It is based on selecting quadruply charged precursors as potentially being cross-linked peptides and then searching the obtained spectra one at a time to try to determine whether they are cross-linked peptides and identify the sequences involved. (
  • Thus, one can identify unmodified and cross-linked peptides together in a single search result. (
  • In all cross-linked protein complexes tested, our new strategy has led to the discovery of not only all cross-linked peptides previously identified through manual analysis but new cross-linked peptides as well. (
  • All chemicals were of analytical reagent grade. (
  • Carbodiimide crosslinking reagents, DCC and EDC (EDAC), for conjugating carboxyl groups (glutamate, aspartate, C-termini) to primary amines (lysine, N-termini) and also N-hydroxysuccinimide (NHS) for stable activation of carboxylates for amine-conjugation. (
  • Upon incubation of detergent-solubilized NADPH-cytochrome P-450 reductase and either cytochrome b5 or cytochrome c in the presence of a water-soluble carbodiimide, a 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC), covalently cross-linked complex was formed. (
  • Alternatively, under conventional methodology, using both BDDGE and 1-Ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS) as cross-linkers, hydrogels were fabricated under acidic conditions. (
  • Nevertheless, we have shown that epoxy cross-linkers should be further explored in the fabrication of collagen-based hydrogels, as alternatives to or in conjunction with carbodiimide cross-linkers. (
  • Carbodiimide cross-linking reagent. (
  • Cross-linkers may be divided into two general categories, homobifunctional and heterobifunctional . (
  • A variety of homobifunctional cross-linkers such as BSOCOES , DSS , DST , Sulfo DST , and DTSSP are available. (
  • Homobifunctional cross-linkers have two identical reactive ends and are used in one-step reactions. (
  • Homobifunctional sulfhydryl-specific crosslinking reagents based on maleimide or pyridyldithiol reactive groups for selective covalent conjugation of protein and peptide thiols (reduced cysteines) to form stable thioether bonds. (
  • Peptide cross-links formed using the homobifunctional-linker diethyl suberthioimidate (DEST) are shown to be ETD-cleavable. (
  • The two functional groups in the crosslinker could be identical as in homobifunctional reagents (Chapter 5), or different as in heterobifunctional reagents (Chapter 6). (
  • The reaction rate for the cross-linking was very high, and for 1 and 3, the reaction reached completion within 1 h (at room temperature). (
  • These reagents are designed to reduce reaction optimization, set-up, and procedure development. (
  • whereas excess TCEP usually does not need to be removed before carrying out the crosslinking reaction. (
  • DSS protein crosslinker must be dissolved in an organic solvent, such as DMSO or DMF , then added to an aqueous crosslinking reaction. (
  • Reducing SDS-polyacrylamide gel electrophoresis analysis of the products from lb indicated the formation of S-ETAC-S interchain heavy and light chain cross-links (~ 23-34% overall yield by video-camera densitometry) which do not undergo disulfide-thiol exchange with DTT at 100 °C. In contrast, no interchain cross-links were observed upon reaction of unreduced or reduced antibody wherein the thiols have been previously alkylated with iodoacetamide. (
  • An efficient iron-catalyzed cross-coupling reaction between alkynyl bromides and Grignard-derived organocuprates has been developed. (
  • During the cross-linking reaction, atoms are eliminated from the reactants, thus shortening the distance between the two linked moieties. (
  • Chitosan/coconut (CTS/coconut) composite membranes w ere successfully prepared by the cross-linking reaction with glutaraldehyde and they were applied in eliminating heavy metals from aqueous solutions. (
  • Chemical cross-linking has been an established method to study protein interaction partners for decades ( 3 ) and has been recently revived by conjunctional mass spectrometric analysis of the cross-linking reaction products ( 4 ). (
  • The cross-linking reaction was carried out in acetic acid for twenty hours at room temperature. (
  • Labeling of the antibody trastuzumab with this reagent provided a labeled antibody conjugate that demonstrated potent HIV-1 neutralization activity demonstrating the potential of this reaction in creating protein conjugates with small molecules. (
  • Collision-induced dissociative chemical cross-linking reagents and methodology: Applications to protein structural characterization using tandem mass spectrometry analysis. (
  • The invention is directed to multi-functional N-maleimidyl polymer derivatives comprising a water soluble and non-peptidic polymer backbone having a terminal carbon, such as a poly(alkylene glycol), the terminal carbon of the polymer backbone being directly bonded to the nitrogen atom of a N-maleimidyl moiety without a linking group therebetween. (
  • 7. A crosslinked polymer formed by the trimerization of an ethynyl radical of a polymer as defined by claim 1. (
  • Surface fabrication methods for patterned deposition of nanometer thick brushes or micron thick, crosslinked films of an azlactone block co-polymer are reported. (
  • While polymer chains in uncross-linked BC/Col are intertwined by H-bonds, new covalent bonds in enzymatically cross-linked ones are formed-resulting in increased thermal stability and crystallinity of the material. (
  • Starch a biodegradable polymer was used in preparation of microspheres using epichlorhydrine as cross-linking agent. (
  • Here, hydrogels are produced via the combination of the protein bovine serum albumin (BSA) and crosslinking reagents, Tetrakis(hydroxymethyl)phosphonium chloride (THPC) and Glutaraldehyde (GA). Shape programing is achieved by creating a temporary secondary crosslinking network by incubating hydrogels in cation solutions (Zn2+ and Cu2+). (
  • Glutaraldehyde was used as a cross-linking reagent for zein (corn protein) to provide fibers with improved physical properties and solvent resistance. (
  • This resulted in a predominant product consisting of two ferulic acid molecules dehydrogenatively linked to a single peptide and, furthermore, two ferulic acids linked to peptide oligomers, ranging from dimers to pentamers. (
  • Protein cross-linking or bioconjugation is the process of creating covalent bonds between two or more molecules. (
  • Crosslinking is the process of chemically joining two or more molecules by a covalent bond. (
  • Upon mAb-mediated cross-linking, p46 molecules induced strong cell triggering leading to [Ca2+]i increases, lymphokine production, and cytolytic activity both in resting NK cells and NK cell clones. (
  • Chemical cross-linking with thiol-cleavable reagents combined with differential mass spectrometric peptide mapping--a novel approach to assess intermolecular protein contacts. (
  • ParR dimers were modified in vitro with the thiol-cleavable cross-linker 3,3'-dithio-bis(succinimidylproprionate) (DTSSP), proteolytically digested with trypsin and analyzed by MALDI-MS peptide mapping. (
  • DXMSMS Match Program for Automated Analysis of LC-MS/MS Data Obtained Using Isotopically Coded CID-Cleavable Cross-Linking Reagents. (
  • Crosslink amines to sulfhydryls with this water-soluble sulfo-NHS-ester and pyridyldithiol groups on 10-atom spacer arms to form cleavable disulfide bonds with cysteines. (
  • In a common workflow with cleavable linkers, characteristic cross-link fragments are observed in MS 2 spectra. (
  • Cross-linkers cleavable by electron transfer dissociation (ETD) have received considerably less attention. (
  • Cross-linkers with ETD-cleavable features might enhance the confidence of cross-link identifications. (
  • However, only a few ETD-cleavable cross-linkers have been reported. (
  • Another ETD-cleavable cross-linker, 1,3-diformyl-5-ethynylbenzene (DEB), forms cross-links through reductive amination. (
  • To facilitate the detection and separation of cross-linked species, various moieties have been introduced into the cross-linking reagents including fluorophores ( 6 ), isotope tags ( 7 , 8 ), cleavable sites ( 9 ), affinity handles ( 10 - 14 ), and a benzyl marker for tandem mass spectrometric analysis ( 15 ). (
  • The ability to synthesis hydrogels and program a reversable temporary shape through three-dimensional crosslinking is essential to the future of medicine and soft robotics. (
  • Other chapters consider the developments in the use of solid support and reagents in peptide synthesis. (
  • 5 . The stent of claim 2 or 3 , further comprising a jacket, a covering or an encapsulation selected from the group consisting of a cross-linked PVA hydrogel, ePTFE, PTFE, porous HDPE, polyurethane, and polyethylene terephthalate. (
  • 6 . The medical device of claim 2 , wherein the synthetic vascular graft comprises a material selected from the group consisting of cross-linked polyvinyl alcohol, ePTFE, PTFE, porous HDPE, polyurethane, and polyethylene terephthalate. (
  • Biotinylate glycoproteins (aldehydes of oxidized sugars) with this short (19.7A) hydrazide-activated biotinylation reagent which also contains a second amine for conjugation. (
  • Chemoselctive ligation refers to the use of mutually specific pairs of conjugation reagents. (
  • Crosslink amines to sulfhydryls with this mid-length, water-soluble crosslinker composed of sulfo-NHS-ester and iodoacetyl (haloacetyl) groups at ends of a 9-atom spacer arm. (
  • Crosslink sulfhydryls to periodate-treated glycoprotein sugars with this long (17.9A) crosslinker containing maleimide and hydrazide groups. (
  • Bruker plans to commercialize the results of the collaboration as integrated solutions for the study of protein structures and interactions using XL-MS. Combining the novel, enrichable PhoX crosslinker 2 , developed by Heck and Scheltema, with the extreme speed and sensitivity of PASEF methods on the timsTOF Pro platform, enables the discovery of more crosslinked products which yield more information about protein structures and interactions. (
  • In our methodology, two transglutaminases are used for enzymatic cross-linking-one recommended for the meat and the other proposed for the fish industry-and pre-oxidated BC (oxBC) is used for chemical cross-linking. (
  • APDP A sulfhydryl-reactive and photoreactive cross-linker Features Chemical Name: N-[4-(p-Azidosalicylamido)butyl]-3'-(2'-pyridyldithio)propionamide Synonym: APDP Formula: C₁₈H₂₀N₆O₂ Molecular Weight: 446.55 Spacer Arm: 21.0Å Reactive Toward: sulfhydryl and amino. (
  • The intermolecular contact regions between monomers of the homodimeric DNA binding protein ParR and the interaction between the glycoproteins CD28 and CD80 were investigated using a strategy that combined chemical cross-linking with differential MALDI-MS analyses. (
  • Here, we developed a novel two-step chemical crosslinking strategy to overcome these problems which resulted in a dramatic improvement in identifying Akt interacting partners. (
  • Chemical cross-linking combined with mass spectrometry is a viable approach to study the low-resolution structure of protein and protein complexes. (
  • Here we describe advances in chemical and computational methods to obtain density of 1 cross-link per 7 amino acids of protein sequence. (
  • Chemical cross-linking coupled with mass spectrometry offers an alternative that has seen increased use, especially in combination with other experimental approaches like cryo-electron microscopy. (
  • Moreover, formaldehyde and other chemical cross-linkers may not enter the cores of large complexes. (
  • The Chemical Cross-linking of Peptide Chains 6. (
  • In this article, we discuss the critical steps of chemical cross-linking and its implications for (structural) biology: reagent design and cross-linking protocols, separation and mass spectrometric analysis of cross-linked samples, dedicated software for data analysis, and the use of cross-linking data for computational modeling. (
  • The concept of protein cross-linking as a (bio)chemical tool to infer structural information about protein conformations and protein-protein interactions in combination with mass spectrometry was introduced at the end of the 1990s ( 1 ). (
  • We critically discuss advantages and limitations of different concepts and look beyond the immediate outcome of cross-linking experiments (putative interactions and/or distance constraints) and examine the potential role of chemical cross-linking in the analysis of protein interaction networks and, more generally, for structural and systems biology. (
  • To date, we have demonstrated that both methods of chemical cross-linking improved the elasticity and tensile strength of the collagen implants. (
  • This article compares the properties of bacterial cellulose/fish collagen composites (BC/Col) after enzymatic and chemical cross-linking. (
  • The mixed disulfide can then be reacted with a reducing agent such as DTT ( D1532 ) or TCEP ( T2556 ) to yield a 3-mercaptopropionyl conjugate or with a thiol-containing biomolecule to form a disulfide-linked tandem conjugate. (
  • Two models of carbohydrate-protein conjugate have been established, the single ended model (terminal amination-single method) and cross-linked lattice matrix (dual amination method). (
  • DSS crosslinking reagent has amine-reactive NHS esters at both ends of an 8-atom (11.4 angstrom) spacer arm. (
  • As products, covalently coupled di- to pentamers of the peptide were identified by LC-MS. Oxidative cross-linking of ferulic acid with horseradish peroxidase and hydrogen peroxide resulted in the formation of dehydrodimers. (
  • For both protein preparations, the aggregates consisted of covalently linked monomers. (
  • The mature particle is formed by 420 copies of the major capsid protein organized on a T = 7 laevo lattice with each subunit covalently crosslinked to two neighbors. (
  • The cross-linking and immunoprecipitation (CLIP) method identified protein-RNA interaction sites in mammalian cells by cloning of the covalently-attached RNAs ( 6 , 7 ). (
  • 10. The compound of claim 9, wherein at least one of Z 1 and Z 2 is --C(═Y)NH-T 3 -Z 3 wherein Z 3 is selected from an iodoacetyl group, a maleimide, a photoreactive linking group, an aldehyde and an epoxide. (
  • From a logical point of view, cross-linking reagents would contain two functional groups, which react with target components, thus linking them together. (
  • it is also possible and, depending on the sample, even more likely that only one end of the bifunctional cross-linker will react with the protein because the other end does not come into contact with another cross-linkable residue, or the second reactive group is deactivated, e.g. by hydrolysis, before forming a cross-link. (
  • These groups react under mild conditions to form disulfide cross‐links by thiol‐disulfide interchange. (
  • A selection of alkylation reagents are offered including 4-Vinylpyridine and OneQuant™ Iodoacetamide . (
  • The site-specific intramolecular cross-linking of sulfhydryls of monoclonal antibodies via a new class of "equilibrium transfer alkylation cross-link (ETAC) reagents" is described. (
  • Additional reagents and equipment for denaturing polyacrylamide gel electrophoresis and UV shadowing ( appendix 3B , CPMB UNIT or Sambrook et al. (
  • Additional reagents and equipment for denaturing polyacrylamide gel electrophoresis ( appendix 3B , CPMB UNIT or Sambrook et al. (
  • A guide to 1D and 2D protein electrophoresis products, including protein markers, electrophoresis buffers, 2D electrophoresis reagents, clean-up reagents and stains. (
  • ADCs containing the DNA alkylator displayed similar in vitro potency, but improved bystander killing and in vivo efficacy, compared with those of the cross-linker. (
  • Cross-linking was successfully performed either in vitro on purified complexes or in vivo in living cells. (
  • Rrp9 Efficiently Cross-Links to the U3 snoRNA in Vitro and in Vivo. (
  • It is noteworthy that 15d-PGJ 2 elicited GSTP1-1 cross-linking in vitro, a process that could be mimicked by other dienone cyclopentenone PG, such as Δ 12 -PGJ 2 , and by the bifunctional thiol reagent dibromobimane, suggesting that cyclopentenone PG may be directly involved in oligomer formation. (
  • Also provided are crosslinked polypeptide compounds and kits that include crosslinking reagents. (
  • In an attempt to improve the clinical success rate of ADCs, emphasis has been recently placed on the use of DNA-cross-linking pyrrolobenzodiazepine compounds as the payload. (
  • These compounds are generally simple and readily available and may be classiŠed into two types: the monofunctional cross-linkers and the zero-length cross-linkers. (
  • Other cross-linking areas such as protein-DNA cross-linking, photoinduced cross-linking, or the characterization of disulfide bonds will not be covered in detail in this paper. (
  • Heterobifunctional cross-linkers have two different reactive ends and are used in two-step sequential reactions. (
  • Primary amine + sulfhydryl reactive heterobifunctional cross-linkers MBS , Sulfo MBS , GMBS , sulfo GMBS , EMCH and others are available. (
  • Also available are a large selection of photoreactive cross-linkers including ABH , APG , NHS-ASA , Sulfo SAND , and many others. (
  • For additional information about protein cross-linkers download our free handbook . (
  • All of these incorporate functional groups that form labile sites along the spacer arms of cross-linkers. (
  • Cisplatin is different from current cross-linking reagents by targeting new functional groups, thioethers, and imidazoles groups, which provides complementarity with existing cross-linkers. (
  • Although monofunctional cross-linkers have one reactive group, part of the molecule is incorporated in the Šnal cross-linked product. (
  • Reagents that catalyze the formation of disulŠde bonds, for example, are zerolength cross-linkers. (
  • A detailed guide to protein cross-linkers and their reactions. (
  • These results indicated site-specific cross-linking of interchain sulfhydryls and places their distance within 3-4 A. Flow cytometry of the ETAC lb 5G6.4 cross-linked product using 77 IP3 human ovarian carcinoma target cells showed positive binding and retention of immunoreactivity. (
  • The present study illustrates the potential applications of labelable ETAC reagents as thiol-specific probes for a wide variety of immunological studies. (
  • Moreover, the unique isotopic pattern of platinum flags cross-linking products and modification sites by mass spectrometry. (
  • Two synthetic strategies were explored in the preparation of the cross-linked collagen scaffolds. (
  • A new synthetic route to reduced collagen crosslinks (LNL and HLNL) is described in this report. (
  • A bi-functional epoxy-based cross-linker, 1,4-Butanediol diglycidyl ether (BDDGE), was investigated in the fabrication of collagen based corneal substitutes. (
  • We employed a new biocompatible crosslinking agent β-cyclodextrin dialdehyde (β-CD-DA) 8 to enhance the mechanical strength of the collagen-based corneal repair material. (
  • The primary amine + carboxyl reactive cross-linker EDC is also offered. (
  • G-Biosciences offer a wide range of tools for protein cross-linking and protein modification applications. (
  • Our high-quality reagents help researchers consistently achieve the protein cross-linking and protein modification results they require. (
  • G-Biosciences provide several protein reduction and modification reagents. (
  • In addition to our protein cross-linking and protein modification reagents, a variety of accessories including SpinOUT™ Columns , Optimizer Buffer™ , Tube-O-Reactor™ , and Tube-O-DIALYZER™ are available to ensure researchers achieve optimal results. (
  • This area of chemistry is known as bioconjugation and includes crosslinking, immobilization, surface modification, and labeling of biomolecules. (
  • Higher charge states not only promote the detection of cross-linking products with less purification, but result in more comprehensive MS/MS fragmentation and can assist the assignment of modification sites. (
  • Another strategy has just been reported where mass modification searching allowed identification of cross-linked candidates ( 20 ). (
  • A recent study with disuccinimidyl suberate (DSS) and bis(sulfosuccinimidyl)suberate (BS3) cross-links showed encouraging results: ETD with supplementary activation of HCD (EThcD) led to the best sequence coverage for highly charged cross-links [ 39 ]. (
  • FGF-2 was cross-linked with bis(sulfosuccinimidyl) suberate, purified by size exclusion chromatography, and digested with trypsin. (
  • Initial design concepts were characterized using a synthesized cross-linked peptide complex. (
  • Following verification and subsequent optimization of cross-linked peptide complex dissociation, our reagents were applied to homodimeric glutathione S-transferase and monomeric bovine serum albumin. (
  • ETD mass pairs can be used in MS 3 experiments to confirm cross-link identifications. (
  • Common LC/MS/MS acquisition approaches such as data-dependent acquisition experiments using ion trap mass spectrometers and product ion spectral analysis using SEQUEST were shown to be compatible with our CID-CXL-MS/MS reagents, obviating the requirement for high resolution and high mass accuracy measurements to identify both intra- and interpeptide cross-links. (
  • Palladium has become one of the most commonly used metal for cross-coupling transformations. (
  • This approach contrasts with other cross-linking reactions in which a spacer is always incorporated between the two cross-linked groups. (
  • A variety of reagents to facilitate this technique are available including the arginine specific endopeptidase SG-Arginine-C™ , the serine endopeptidase SG-Chymotrypsin™ , the serine endopeptidase SG-Glutamic-C™ , and the highly specific serine protease SG-Lysine-C™ . Also available is the chemically methylated, TPCK treated affinity purified Mass Spectrometry Grade Trypsin . (
  • In addition, platinum(II) inherently has two positive charges which enhance the detection of cross-linked products. (
  • Albert Heck of Utrecht University commented: "We are delighted to work with Bruker on the further development of workflows for XL-MS that take advantage of the speed of PASEF and the unique large-scale, accurate CCS data to enhance the detection of crosslinks in XL-MS. We are excited by the initial results published in Molecular and Cellular Proteomics and look forward to advancing XL-MS even further. (
  • Both reagents act as a bridge for crosslinking between BSA amino acid binding sites. (
  • Other reagents condense carboxyl and primary amino groups to form amide bonds, hydroxyl and carboxyl groups to form esters, thiol and carboxyl groups to form thioesters, and so on. (
  • Many of these reagents simply act as activating agents converting one of the components, for example, carboxyl groups, into a reactive species. (
  • These features provide a simple but reliable approach to identify cross-links that should facilitate studies of protein complexes. (
  • Recent advances in instrumentation, cross-linking protocols, and analysis software have led to a renewed interest in this technique, which promises to provide important information about native protein structure and the topology of protein complexes. (