Chromatography, High Pressure Liquid: Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.Chromatography, Affinity: A chromatographic technique that utilizes the ability of biological molecules to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)SepharoseChromatography, Gel: Chromatography on non-ionic gels without regard to the mechanism of solute discrimination.Chromatography: Techniques used to separate mixtures of substances based on differences in the relative affinities of the substances for mobile and stationary phases. A mobile phase (fluid or gas) passes through a column containing a stationary phase of porous solid or liquid coated on a solid support. Usage is both analytical for small amounts and preparative for bulk amounts.Chromatography, Ion Exchange: Separation technique in which the stationary phase consists of ion exchange resins. The resins contain loosely held small ions that easily exchange places with other small ions of like charge present in solutions washed over the resins.Chromatography, Gas: Fractionation of a vaporized sample as a consequence of partition between a mobile gaseous phase and a stationary phase held in a column. Two types are gas-solid chromatography, where the fixed phase is a solid, and gas-liquid, in which the stationary phase is a nonvolatile liquid supported on an inert solid matrix.Electrophoresis, Agar Gel: Electrophoresis in which agar or agarose gel is used as the diffusion medium.Chromatography, Liquid: Chromatographic techniques in which the mobile phase is a liquid.Chromatography, Thin Layer: Chromatography on thin layers of adsorbents rather than in columns. The adsorbent can be alumina, silica gel, silicates, charcoals, or cellulose. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Chromatography, Agarose: A method of gel filtration chromatography using agarose, the non-ionic component of agar, for the separation of compounds with molecular weights up to several million.Molecular Weight: The sum of the weight of all the atoms in a molecule.Electrophoresis, Polyacrylamide Gel: Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.Chromatography, DEAE-Cellulose: A type of ion exchange chromatography using diethylaminoethyl cellulose (DEAE-CELLULOSE) as a positively charged resin. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Molecular Sequence Data: 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.Kinetics: The rate dynamics in chemical or physical systems.Mass Spectrometry: An analytical method used in determining the identity of a chemical based on its mass using mass analyzers/mass spectrometers.Amino Acid Sequence: 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.Gas Chromatography-Mass Spectrometry: A microanalytical technique combining mass spectrometry and gas chromatography for the qualitative as well as quantitative determinations of compounds.Amino Acids: 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.Chromatography, Reverse-Phase: A chromatography technique in which the stationary phase is composed of a non-polar substance with a polar mobile phase, in contrast to normal-phase chromatography in which the stationary phase is a polar substance with a non-polar mobile phase.Chromatography, Paper: An analytical technique for resolution of a chemical mixture into its component compounds. Compounds are separated on an adsorbent paper (stationary phase) by their varied degree of solubility/mobility in the eluting solvent (mobile phase).Isoelectric Focusing: Electrophoresis in which a pH gradient is established in a gel medium and proteins migrate until they reach the site (or focus) at which the pH is equal to their isoelectric point.Cattle: 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.Hydrogen-Ion Concentration: 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)Chemistry: A basic science concerned with the composition, structure, and properties of matter; and the reactions that occur between substances and the associated energy exchange.Chemical Phenomena: The composition, conformation, and properties of atoms and molecules, and their reaction and interaction processes.Substrate Specificity: A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.Escherichia coli: 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.Immunodiffusion: Technique involving the diffusion of antigen or antibody through a semisolid medium, usually agar or agarose gel, with the result being a precipitin reaction.Methods: A series of steps taken in order to conduct research.Electrophoresis: An electrochemical process in which macromolecules or colloidal particles with a net electric charge migrate in a solution under the influence of an electric current.Spectrophotometry, Ultraviolet: 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)Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Carbohydrates: The largest class of organic compounds, including STARCH; GLYCOGEN; CELLULOSE; POLYSACCHARIDES; and simple MONOSACCHARIDES. Carbohydrates are composed of carbon, hydrogen, and oxygen in a ratio of Cn(H2O)n.Immunoelectrophoresis: A technique that combines protein electrophoresis and double immunodiffusion. In this procedure proteins are first separated by gel electrophoresis (usually agarose), then made visible by immunodiffusion of specific antibodies. A distinct elliptical precipitin arc results for each protein detectable by the antisera.Tandem Mass Spectrometry: A mass spectrometry technique using two (MS/MS) or more mass analyzers. With two in tandem, the precursor ions are mass-selected by a first mass analyzer, and focused into a collision region where they are then fragmented into product ions which are then characterized by a second mass analyzer. A variety of techniques are used to separate the compounds, ionize them, and introduce them to the first mass analyzer. For example, for in GC-MS/MS, GAS CHROMATOGRAPHY-MASS SPECTROMETRY is involved in separating relatively small compounds by GAS CHROMATOGRAPHY prior to injecting them into an ionization chamber for the mass selection.Isoelectric Point: The pH in solutions of proteins and related compounds at which the dipolar ions are at a maximum.DNA: 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).Rabbits: 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.Chemical Fractionation: Separation of a mixture in successive stages, each stage removing from the mixture some proportion of one of the substances, for example by differential solubility in water-solvent mixtures. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)PolysaccharidesMacromolecular Substances: 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.Protein Binding: 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.Spectrometry, Mass, Electrospray Ionization: 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.Drug Stability: The chemical and physical integrity of a pharmaceutical product.Indicators and Reagents: 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)Temperature: 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.Liver: A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.Carbohydrate Sequence: The sequence of carbohydrates within POLYSACCHARIDES; GLYCOPROTEINS; and GLYCOLIPIDS.Peptide Fragments: Partial proteins formed by partial hydrolysis of complete proteins or generated through PROTEIN ENGINEERING techniques.Oligosaccharides: Carbohydrates consisting of between two (DISACCHARIDES) and ten MONOSACCHARIDES connected by either an alpha- or beta-glycosidic link. They are found throughout nature in both the free and bound form.Gels: Colloids with a solid continuous phase and liquid as the dispersed phase; gels may be unstable when, due to temperature or other cause, the solid phase liquefies; the resulting colloid is called a sol.Recombinant Proteins: Proteins prepared by recombinant DNA technology.Countercurrent Distribution: A method of separation of two or more substances by repeated distribution between two immiscible liquid phases that move past each other in opposite directions. It is a form of liquid-liquid chromatography. (Stedman, 25th ed)Trypsin: 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 3.4.21.4.Chromatography, Micellar Electrokinetic Capillary: A hybrid separation technique combining both chromatographic and electrophoretic separation principles. While the method was invented to separate neutral species, it can also be applied to charged molecules such as small peptides.Solubility: The ability of a substance to be dissolved, i.e. to form a solution with another substance. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)Hydrolysis: The process of cleaving a chemical compound by the addition of a molecule of water.Magnetic Resonance Spectroscopy: 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).Centrifugation, Density Gradient: Separation of particles according to density by employing a gradient of varying densities. At equilibrium each particle settles in the gradient at a point equal to its density. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Cloning, Molecular: The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.Glycoproteins: Conjugated protein-carbohydrate compounds including mucins, mucoid, and amyloid glycoproteins.Chemical Precipitation: The formation of a solid in a solution as a result of a chemical reaction or the aggregation of soluble substances into complexes large enough to fall out of solution.Cell Line: Established cell cultures that have the potential to propagate indefinitely.Peptides: 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.Plasmids: Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS.Ultracentrifugation: Centrifugation with a centrifuge that develops centrifugal fields of more than 100,000 times gravity. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Glycoside HydrolasesHydroxyapatites: A group of compounds with the general formula M10(PO4)6(OH)2, where M is barium, strontium, or calcium. The compounds are the principal mineral in phosphorite deposits, biological tissue, human bones, and teeth. They are also used as an anticaking agent and polymer catalysts. (Grant & Hackh's Chemical Dictionary, 5th ed)Reference Standards: 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.Cells, Cultured: 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.Isoenzymes: Structurally related forms of an enzyme. Each isoenzyme has the same mechanism and classification, but differs in its chemical, physical, or immunological characteristics.Species Specificity: 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.Reproducibility of Results: 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.Polymerase Chain Reaction: 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.Binding Sites: The parts of a macromolecule that directly participate in its specific combination with another molecule.Swine: 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).Microchemistry: 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.Sensitivity and Specificity: 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)DNA, Bacterial: Deoxyribonucleic acid that makes up the genetic material of bacteria.Radioimmunoassay: 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.Glycosaminoglycans: Heteropolysaccharides which contain an N-acetylated hexosamine in a characteristic repeating disaccharide unit. The repeating structure of each disaccharide involves alternate 1,4- and 1,3-linkages consisting of either N-acetylglucosamine or N-acetylgalactosamine.Immunoelectrophoresis, Two-Dimensional: Immunoelectrophoresis in which a second electrophoretic transport is performed on the initially separated antigen fragments into an antibody-containing medium in a direction perpendicular to the first electrophoresis.Culture Media: Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN.Lectins: Proteins that share the common characteristic of binding to carbohydrates. Some ANTIBODIES and carbohydrate-metabolizing proteins (ENZYMES) also bind to carbohydrates, however they are not considered lectins. PLANT LECTINS are carbohydrate-binding proteins that have been primarily identified by their hemagglutinating activity (HEMAGGLUTININS). However, a variety of lectins occur in animal species where they serve diverse array of functions through specific carbohydrate recognition.Carbohydrate Conformation: The characteristic 3-dimensional shape of a carbohydrate.DNA Restriction Enzymes: Enzymes that are part of the restriction-modification systems. They catalyze the endonucleolytic cleavage of DNA sequences which lack the species-specific methylation pattern in the host cell's DNA. Cleavage yields random or specific double-stranded fragments with terminal 5'-phosphates. The function of restriction enzymes is to destroy any foreign DNA that invades the host cell. Most have been studied in bacterial systems, but a few have been found in eukaryotic organisms. They are also used as tools for the systematic dissection and mapping of chromosomes, in the determination of base sequences of DNAs, and have made it possible to splice and recombine genes from one organism into the genome of another. EC 3.21.1.Time Factors: Elements of limited time intervals, contributing to particular results or situations.Bacterial Proteins: Proteins found in any species of bacterium.Hot Temperature: Presence of warmth or heat or a temperature notably higher than an accustomed norm.Biological Assay: A method of measuring the effects of a biologically active substance using an intermediate in vivo or in vitro tissue or cell model under controlled conditions. It includes virulence studies in animal fetuses in utero, mouse convulsion bioassay of insulin, quantitation of tumor-initiator systems in mouse skin, calculation of potentiating effects of a hormonal factor in an isolated strip of contracting stomach muscle, etc.Nucleic Acid Hybridization: Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503)Molecular Structure: 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.Calibration: 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.Microscopy, Electron: 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.Blood Proteins: Proteins that are present in blood serum, including SERUM ALBUMIN; BLOOD COAGULATION FACTORS; and many other types of proteins.Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization: A mass spectrometric technique that is used for the analysis of large biomolecules. Analyte molecules are embedded in an excess matrix of small organic molecules that show a high resonant absorption at the laser wavelength used. The matrix absorbs the laser energy, thus inducing a soft disintegration of the sample-matrix mixture into free (gas phase) matrix and analyte molecules and molecular ions. In general, only molecular ions of the analyte molecules are produced, and almost no fragmentation occurs. This makes the method well suited for molecular weight determinations and mixture analysis.Carrier Proteins: Transport proteins that carry specific substances in the blood or across cell membranes.Rats, Inbred Strains: Genetically identical individuals developed from brother and sister matings which have been carried out for twenty or more generations or by parent x offspring matings carried out with certain restrictions. This also includes animals with a long history of closed colony breeding.Cyanogen Bromide: Cyanogen bromide (CNBr). A compound used in molecular biology to digest some proteins and as a coupling reagent for phosphoroamidate or pyrophosphate internucleotide bonds in DNA duplexes.Proteins: 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.Electrophoresis, Cellulose Acetate: Electrophoresis in which cellulose acetate is the diffusion medium.Fatty Acids: Organic, monobasic acids derived from hydrocarbons by the equivalent of oxidation of a methyl group to an alcohol, aldehyde, and then acid. Fatty acids are saturated and unsaturated (FATTY ACIDS, UNSATURATED). (Grant & Hackh's Chemical Dictionary, 5th ed)Acetonitriles: Compounds in which a methyl group is attached to the cyano moiety.Electrophoresis, Gel, Two-Dimensional: Electrophoresis in which a second perpendicular electrophoretic transport is performed on the separate components resulting from the first electrophoresis. This technique is usually performed on polyacrylamide gels.Sequence Homology, Amino Acid: The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.Brain Chemistry: Changes in the amounts of various chemicals (neurotransmitters, receptors, enzymes, and other metabolites) specific to the area of the central nervous system contained within the head. These are monitored over time, during sensory stimulation, or under different disease states.Plant Extracts: Concentrated pharmaceutical preparations of plants obtained by removing active constituents with a suitable solvent, which is evaporated away, and adjusting the residue to a prescribed standard.Detergents: Purifying or cleansing agents, usually salts of long-chain aliphatic bases or acids, that exert cleansing (oil-dissolving) and antimicrobial effects through a surface action that depends on possessing both hydrophilic and hydrophobic properties.Stereoisomerism: 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)Densitometry: The measurement of the density of a material by measuring the amount of light or radiation passing through (or absorbed by) the material.Phospholipids: Lipids containing one or more phosphate groups, particularly those derived from either glycerol (phosphoglycerides see GLYCEROPHOSPHOLIPIDS) or sphingosine (SPHINGOLIPIDS). They are polar lipids that are of great importance for the structure and function of cell membranes and are the most abundant of membrane lipids, although not stored in large amounts in the system.DNA Fragmentation: Splitting the DNA into shorter pieces by endonucleolytic DNA CLEAVAGE at multiple sites. It includes the internucleosomal DNA fragmentation, which along with chromatin condensation, are considered to be the hallmarks of APOPTOSIS.Enzyme Stability: The extent to which an enzyme retains its structural conformation or its activity when subjected to storage, isolation, and purification or various other physical or chemical manipulations, including proteolytic enzymes and heat.Ammonium Sulfate: Sulfuric acid diammonium salt. It is used in CHEMICAL FRACTIONATION of proteins.Proteoglycans: Glycoproteins which have a very high polysaccharide content.DNA Primers: 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.Antibodies, Monoclonal: Antibodies produced by a single clone of cells.Durapatite: The mineral component of bones and teeth; it has been used therapeutically as a prosthetic aid and in the prevention and treatment of osteoporosis.Oxidation-Reduction: 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).Chickens: Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA.Disaccharides: Oligosaccharides containing two monosaccharide units linked by a glycosidic bond.Cell Membrane: The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.TritiumPeptide Mapping: Analysis of PEPTIDES that are generated from the digestion or fragmentation of a protein or mixture of PROTEINS, by ELECTROPHORESIS; CHROMATOGRAPHY; or MASS SPECTROMETRY. The resulting peptide fingerprints are analyzed for a variety of purposes including the identification of the proteins in a sample, GENETIC POLYMORPHISMS, patterns of gene expression, and patterns diagnostic for diseases.Spectrometry, Fluorescence: Measurement of the intensity and quality of fluorescence.Blood Protein Electrophoresis: Electrophoresis applied to BLOOD PROTEINS.Limit of Detection: Concentration or quantity that is derived from the smallest measure that can be detected with reasonable certainty for a given analytical procedure.Immune Sera: 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.Solid Phase Extraction: An extraction method that separates analytes using a solid phase and a liquid phase. It is used for preparative sample cleanup before analysis by CHROMATOGRAPHY and other analytical methods.Physicochemical Phenomena: The physical phenomena describing the structure and properties of atoms and molecules, and their reaction and interaction processes.Monosaccharides: Simple sugars, carbohydrates which cannot be decomposed by hydrolysis. They are colorless crystalline substances with a sweet taste and have the same general formula CnH2nOn. (From Dorland, 28th ed)Anion Exchange Resins: High-molecular-weight insoluble polymers that contain functional cationic groups capable of undergoing exchange reactions with anions.Spectrophotometry: The art or process of comparing photometrically the relative intensities of the light in different parts of the spectrum.Binding, Competitive: The interaction of two or more substrates or ligands with the same binding site. The displacement of one by the other is used in quantitative and selective affinity measurements.Chemistry Techniques, Analytical: Methodologies used for the isolation, identification, detection, and quantitation of chemical substances.DNA, Viral: Deoxyribonucleic acid that makes up the genetic material of viruses.Blotting, Western: 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.Methanol: A colorless, flammable liquid used in the manufacture of FORMALDEHYDE and ACETIC ACID, in chemical synthesis, antifreeze, and as a solvent. Ingestion of methanol is toxic and may cause blindness.Ultrafiltration: The separation of particles from a suspension by passage through a filter with very fine pores. In ultrafiltration the separation is accomplished by convective transport; in DIALYSIS separation relies instead upon differential diffusion. Ultrafiltration occurs naturally and is a laboratory procedure. Artificial ultrafiltration of the blood is referred to as HEMOFILTRATION or HEMODIAFILTRATION (if combined with HEMODIALYSIS).Chemistry, Physical: The study of CHEMICAL PHENOMENA and processes in terms of the underlying PHYSICAL PHENOMENA and processes.Polymers: Compounds formed by the joining of smaller, usually repeating, units linked by covalent bonds. These compounds often form large macromolecules (e.g., BIOPOLYMERS; PLASTICS).Solvents: Liquids that dissolve other substances (solutes), generally solids, without any change in chemical composition, as, water containing sugar. (Grant & Hackh's Chemical Dictionary, 5th ed)Sulfates: Inorganic salts of sulfuric acid.Structure-Activity Relationship: 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.Glycopeptides: Proteins which contain carbohydrate groups attached covalently to the polypeptide chain. The protein moiety is the predominant group with the carbohydrate making up only a small percentage of the total weight.Cytosol: Intracellular fluid from the cytoplasm after removal of ORGANELLES and other insoluble cytoplasmic components.Flame Ionization: Pyrolysis of organic compounds at the temperature of a hydrogen-air flame to produce ionic intermediates which can be collected and the resulting ion current measured by gas chromatography.GlucosamineProtein Conformation: 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).Edetic Acid: A chelating agent that sequesters a variety of polyvalent cations such as CALCIUM. It is used in pharmaceutical manufacturing and as a food additive.Immunosorbent Techniques: Techniques for removal by adsorption and subsequent elution of a specific antibody or antigen using an immunosorbent containing the homologous antigen or antibody.Cross Reactions: Serological reactions in which an antiserum against one antigen reacts with a non-identical but closely related antigen.DNA, Circular: Any of the covalently closed DNA molecules found in bacteria, many viruses, mitochondria, plastids, and plasmids. Small, polydisperse circular DNA's have also been observed in a number of eukaryotic organisms and are suggested to have homology with chromosomal DNA and the capacity to be inserted into, and excised from, chromosomal DNA. It is a fragment of DNA formed by a process of looping out and deletion, containing a constant region of the mu heavy chain and the 3'-part of the mu switch region. Circular DNA is a normal product of rearrangement among gene segments encoding the variable regions of immunoglobulin light and heavy chains, as well as the T-cell receptor. (Riger et al., Glossary of Genetics, 5th ed & Segen, Dictionary of Modern Medicine, 1992)Adsorption: The adhesion of gases, liquids, or dissolved solids onto a surface. It includes adsorptive phenomena of bacteria and viruses onto surfaces as well. ABSORPTION into the substance may follow but not necessarily.Galactose: An aldohexose that occurs naturally in the D-form in lactose, cerebrosides, gangliosides, and mucoproteins. Deficiency of galactosyl-1-phosphate uridyltransferase (GALACTOSE-1-PHOSPHATE URIDYL-TRANSFERASE DEFICIENCY DISEASE) causes an error in galactose metabolism called GALACTOSEMIA, resulting in elevations of galactose in the blood.Immunoassay: 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.Cations, Divalent: Positively charged atoms, radicals or groups of atoms with a valence of plus 2, which travel to the cathode or negative pole during electrolysis.Mutation: 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.Evaluation Studies as Topic: 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.Erythrocytes: Red blood cells. Mature erythrocytes are non-nucleated, biconcave disks containing HEMOGLOBIN whose function is to transport OXYGEN.Chymotrypsin: A serine endopeptidase secreted by the pancreas as its zymogen, CHYMOTRYPSINOGEN and carried in the pancreatic juice to the duodenum where it is activated by TRYPSIN. It selectively cleaves aromatic amino acids on the carboxyl side.Heparin: A highly acidic mucopolysaccharide formed of equal parts of sulfated D-glucosamine and D-glucuronic acid with sulfaminic bridges. The molecular weight ranges from six to twenty thousand. Heparin occurs in and is obtained from liver, lung, mast cells, etc., of vertebrates. Its function is unknown, but it is used to prevent blood clotting in vivo and vitro, in the form of many different salts.DNA, Superhelical: Circular duplex DNA isolated from viruses, bacteria and mitochondria in supercoiled or supertwisted form. This superhelical DNA is endowed with free energy. During transcription, the magnitude of RNA initiation is proportional to the DNA superhelicity.Plants: Multicellular, eukaryotic life forms of kingdom Plantae (sensu lato), comprising the VIRIDIPLANTAE; RHODOPHYTA; and GLAUCOPHYTA; all of which acquired chloroplasts by direct endosymbiosis of CYANOBACTERIA. They are characterized by a mainly photosynthetic mode of nutrition; essentially unlimited growth at localized regions of cell divisions (MERISTEMS); cellulose within cells providing rigidity; the absence of organs of locomotion; absence of nervous and sensory systems; and an alternation of haploid and diploid generations.Electrophoresis, Capillary: A highly-sensitive (in the picomolar range, which is 10,000-fold more sensitive than conventional electrophoresis) and efficient technique that allows separation of PROTEINS; NUCLEIC ACIDS; and CARBOHYDRATES. (Segen, Dictionary of Modern Medicine, 1992)Spectrometry, Mass, Fast Atom Bombardment: A mass spectrometric technique that is used for the analysis of a wide range of biomolecules, such as glycoalkaloids, glycoproteins, polysaccharides, and peptides. Positive and negative fast atom bombardment spectra are recorded on a mass spectrometer fitted with an atom gun with xenon as the customary beam. The mass spectra obtained contain molecular weight recognition as well as sequence information.Isomerism: The phenomenon whereby certain chemical compounds have structures that are different although the compounds possess the same elemental composition. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 5th ed)RNA, Messenger: RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm.Polyethylene Glycols: Polymers of ETHYLENE OXIDE and water, and their ethers. They vary in consistency from liquid to solid depending on the molecular weight indicated by a number following the name. They are used as SURFACTANTS, dispersing agents, solvents, ointment and suppository bases, vehicles, and tablet excipients. Some specific groups are NONOXYNOLS, OCTOXYNOLS, and POLOXAMERS.Glycolipids: Any compound containing one or more monosaccharide residues bound by a glycosidic linkage to a hydrophobic moiety such as an acylglycerol (see GLYCERIDES), a sphingoid, a ceramide (CERAMIDES) (N-acylsphingoid) or a prenyl phosphate. (From IUPAC's webpage)Sodium Dodecyl Sulfate: An anionic surfactant, usually a mixture of sodium alkyl sulfates, mainly the lauryl; lowers surface tension of aqueous solutions; used as fat emulsifier, wetting agent, detergent in cosmetics, pharmaceuticals and toothpastes; also as research tool in protein biochemistry.Endopeptidases: A subclass of PEPTIDE HYDROLASES that catalyze the internal cleavage of PEPTIDES or PROTEINS.Antibodies: 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).Plants, Medicinal: Plants whose roots, leaves, seeds, bark, or other constituent parts possess therapeutic, tonic, purgative, curative or other pharmacologic attributes, when administered to man or animals.Electrochemistry: The study of chemical changes resulting from electrical action and electrical activity resulting from chemical changes.HexosaminesChromatography, Supercritical Fluid: A CHROMATOGRAPHY method using supercritical fluid, usually carbon dioxide under very high pressure (around 73 atmospheres or 1070 psi at room temperature) as the mobile phase. Other solvents are sometimes added as modifiers. This is used both for analytical (SFC) and extraction (SFE) purposes.Peptide Hydrolases: Hydrolases that specifically cleave the peptide bonds found in PROTEINS and PEPTIDES. Examples of sub-subclasses for this group include EXOPEPTIDASES and ENDOPEPTIDASES.Sulfhydryl Compounds: Compounds containing the -SH radical.Osmolar Concentration: The concentration of osmotically active particles in solution expressed in terms of osmoles of solute per liter of solution. Osmolality is expressed in terms of osmoles of solute per kilogram of solvent.Immunoblotting: Immunologic method used for detecting or quantifying immunoreactive substances. The substance is identified by first immobilizing it by blotting onto a membrane and then tagging it with labeled antibodies.Biotransformation: The chemical alteration of an exogenous substance by or in a biological system. The alteration may inactivate the compound or it may result in the production of an active metabolite of an inactive parent compound. The alterations may be divided into METABOLIC DETOXICATION, PHASE I and METABOLIC DETOXICATION, PHASE II.Dose-Response Relationship, Drug: The relationship between the dose of an administered drug and the response of the organism to the drug.Autoanalysis: Method of analyzing chemicals using automation.Protease Inhibitors: Compounds which inhibit or antagonize biosynthesis or actions of proteases (ENDOPEPTIDASES).Methylation: Addition of methyl groups. In histo-chemistry methylation is used to esterify carboxyl groups and remove sulfate groups by treating tissue sections with hot methanol in the presence of hydrochloric acid. (From Stedman, 25th ed)Collagen: A polypeptide substance comprising about one third of the total protein in mammalian organisms. It is the main constituent of SKIN; CONNECTIVE TISSUE; and the organic substance of bones (BONE AND BONES) and teeth (TOOTH).Dextrans: A group of glucose polymers made by certain bacteria. Dextrans are used therapeutically as plasma volume expanders and anticoagulants. They are also commonly used in biological experimentation and in industry for a wide variety of purposes.Cell-Free System: A fractionated cell extract that maintains a biological function. A subcellular fraction isolated by ultracentrifugation or other separation techniques must first be isolated so that a process can be studied free from all of the complex side reactions that occur in a cell. The cell-free system is therefore widely used in cell biology. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p166)Uronic Acids: Acids derived from monosaccharides by the oxidation of the terminal (-CH2OH) group farthest removed from the carbonyl group to a (-COOH) group. (From Stedmans, 26th ed)Circular Dichroism: A change from planar to elliptic polarization when an initially plane-polarized light wave traverses an optically active medium. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Enzyme-Linked Immunosorbent Assay: 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.Lipoproteins: Lipid-protein complexes involved in the transportation and metabolism of lipids in the body. They are spherical particles consisting of a hydrophobic core of TRIGLYCERIDES and CHOLESTEROL ESTERS surrounded by a layer of hydrophilic free CHOLESTEROL; PHOSPHOLIPIDS; and APOLIPOPROTEINS. Lipoproteins are classified by their varying buoyant density and sizes.Immunochemistry: Field of chemistry that pertains to immunological phenomena and the study of chemical reactions related to antigen stimulation of tissues. It includes physicochemical interactions between antigens and antibodies.Glycosylation: The chemical or biochemical addition of carbohydrate or glycosyl groups to other chemicals, especially peptides or proteins. Glycosyl transferases are used in this biochemical reaction.Tissue Distribution: Accumulation of a drug or chemical substance in various organs (including those not relevant to its pharmacologic or therapeutic action). This distribution depends on the blood flow or perfusion rate of the organ, the ability of the drug to penetrate organ membranes, tissue specificity, protein binding. The distribution is usually expressed as tissue to plasma ratios.Affinity Labels: 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.Cell Fractionation: Techniques to partition various components of the cell into SUBCELLULAR FRACTIONS.Esters

Globular domains of agrin are functional units that collaborate to induce acetylcholine receptor clustering. (1/422)

Agrin, an extracellular matrix protein involved in neuromuscular junction formation, directs clustering of postsynaptic molecules, including acetylcholine receptors (AChRs). This activity resides entirely in the C-terminal portion of the protein, which consists of three laminin-like globular domains (G-domains: G1, G2 and G3) and four EGF-like repeats. Additionally, alternate mRNA splicing yields G-domain variants G2(0,4) with 0- or 4-amino-acid inserts, and G3(0, 8,11,19) with 0-, 8-, 11- or 19-amino-acid inserts. In order to better understand the contributions of individual domains and alternate splicing to agrin activity, single G-domains and covalently linked pairs of G-domains were expressed as soluble proteins and their AChR clustering activity measured on cultured C2 myotubes. These analyses reveal the following: (1) While only G3(8) exhibits detectable activity by itself, all G-domains studied (G1, G2(0), G2(4), G3(0) and G3(8)) enhance G3(8) activity when physically linked to G3(8). This effect is most pronounced when G2(4) is linked to G3(8) and is independent of the order of the G-domains. (2) The deletion of EGF-like repeats enhances activity. (3) Increasing the physical separation between linked G1 and G3(8) domains produces a significant increase in activity; similar alterations to linked G2 and G3(8) domains are without effect. (4) Clusters induced by two concatenated G3(8) domains are significantly smaller than all other agrin forms studied. These data suggest that agrin G-domains are the functional units which interact independently of their specific organization to yield AChR clustering. G-domain synergism resulting in biological output could be due to physical interactions between G-domains or, alternatively, independent interactions of G-domains with cell surface receptors which require spatially localized coactivation for optimal signal transduction.  (+info)

Pseudomonas ribosomal vaccines: preparation, properties, and immunogenicity. (2/422)

The preparation, properties, and immunogenicity of ribosomal vaccines from Pseudomonas aeruginosa are described. These preparations, containing protein and RNA, were tested for immunogenicity by active immunization of mice and subsequent challenge with homologous, live bacteria. The results demonstrated that vaccines prepared from a majority of serotypes used were immunogenic, i.e., afforded 60 to 100% mouse protection against a challenge inoculum containing 8 to 50 50% lethal doses. In some cases vaccine doses as low as 1 microgram of RNA provided 100% mouse protection. Molecular sieve chromatography of a highly immunogenic ribosomal preparation on Sepharose 4B demonstrated the presence of two molecular weight fractions: (i) peak A, an excluded peak (thus having a molecular weight of at least 2 times 10(7)), and (ii) peak B, considerably retarded, with an elution position corresponding to a molecular weight of about 2.2 X 10(6), approximating that of typical 70S ribosomes. Both peaks A and B were immunogenic; however, the immunogenicity of peak A was greater (i.e., a smaller immunizing dose was required) than that of peak B. Peak A was shown to contain components of lipopolysaccharide in addition to protein and RNA (which comprised 80% of the dry weight of peak A). On the other hand, peak B was shown to be free of lipopolysaccharide, and 100% of its dry weight consisted of protein and RNA.  (+info)

Isolation and characterization of linear polylactosamines containing one and two site-specifically positioned Lewis x determinants: WGA agarose chromatography in fractionation of mixtures generated by random, partial enzymatic alpha3-fucosylation of pure polylactosamines. (3/422)

We report that isomeric monofucosylhexasaccharides, Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1- 3Galbeta1-4(Fucalpha1-3) GlcNAc, Galbeta1-4GlcNAcbeta1-3Galbeta1-4(Fucalpha1-3) GlcNAcbeta1-3Galbeta1-4 GlcNAc and Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-3Galbeta1- 4GlcNAcbeta1-3Galbeta1-4 GlcNAc, and bifucosylhexasaccharides Galbeta1-4GlcNAcbeta1-3Galbeta1-4(Fucalpha1-3) GlcNAcbeta1-3Galbeta1-4(Fucalpha1-3)GlcNAc, Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-3Galbeta1- 4GlcNAcbeta1-3Galbeta1-4 (Fucalpha1-3)GlcNAc and Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-3Galbeta1-4( Fucalpha1-3)GlcNAcbeta1-3Galbeta1-4GlcNAc can be isolated in pure form from reaction mixtures of the linear hexasaccharide Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1- 3Galbeta1-4GlcNAc with GDP-fucose and alpha1,3-fucosyltransferases of human milk. The pure isomers were characterized in several ways;1H-NMR spectroscopy, for instance, revealed distinct resonances associated with the Lewis x group [Galbeta1-4(Fucalpha1-3)GlcNAc] located at the proximal, middle, and distal positions of the polylactosamine chain. Chromatography on immobilized wheat germ agglutinin was crucial in the separation process used; the isomers carrying the fucose at the reducing end GlcNAc possessed particularly low affinities for the lectin. Isomeric monofucosyl derivatives of the pentasaccharides GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1-3Galbeta1- 4Gl cNAc and Galalpha1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4G lcN Ac and the tetrasaccharide Galbeta1-4GlcNAcbeta1-3Galbeta1-4GlcNAc were also obtained in pure form, implying that the methods used are widely applicable. The isomeric Lewis x glycans proved to be recognized in highly variable binding modes by polylactosamine-metabolizing enzymes, e.g., the midchain beta1,6-GlcNAc transferase (Leppanen et al., Biochemistry, 36, 13729-13735, 1997).  (+info)

IgA interaction with carboxy-terminal 43-kD fragment of fibronectin in IgA nephropathy. (4/422)

IgA deposition in the glomerular mesangial matrix is a prerequisite for the diagnosis of IgA nephropathy, and circulating IgA-containing complex has been implicated in this process. Since fibronectin is known to be involved in the assembly of extracellular matrix, this study was conducted to investigate whether fibronectin and its fragments are present in sera of patients and are capable of binding IgA1. Sera from patients with IgA nephropathy were purified by heparin-affinity chromatography, and column eluate were analyzed for the presence of fibronectin using Western blot and a set of anti-fibronectin monoclonal antibodies. Native fibronectin was digested with cathepsin D to obtain fragments similar to those of serum fibronectin. The capacity of fibronectin to bind IgA was examined with a mixture of purified IgA1 and cathepsin D-digested fibronectin fragments. A 43-kD carboxy-terminal fragment of fibronectin was detected in samples derived from sera of patients with IgA nephropathy but not in healthy control subjects. A similar-sized fragment was generated by cathepsin D digestion of the native molecule and was shown to bind to IgA1 in vitro. Since the carboxy-terminal domain is known to be critical in assembling exogenous fibronectin into the extracellular matrix, the affinity to IgA1 to a fragment found in patients may have pathogenic potential to mediate extracellular IgA deposition in IgA nephropathy.  (+info)

Chimeras of human extracellular and intracellular superoxide dismutases. Analysis of structure and function of the individual domains. (5/422)

Human extracellular superoxide dismutase (hEC-SOD) is a secreted tetrameric protein involved in protection against oxygen free radicals. Since EC-SOD is too large a protein for structural determination by multi-dimensional NMR and attempts to crystallize the protein for X-ray structural determination have failed, the three-dimensional structure of hEC-SOD is unknown. By fusion protein techniques we have previously shown that an amphipathic alpha-helix in the N-terminal domain of hEC-SOD is essential for the tetramer interaction. However, the central domain, which is homologous to intracellular hCuZnSOD, has also been proposed to be involved in the tetramer formation. Despite great efforts, the production of recombinant hEC-SOD in prokaryotic systems or simple eukaryotes (such as yeast) has failed. This lack of success has greatly complicated large-scale production and genetic engineering of the protein. In the study reported here, we constructed two chimeras comprising the N- or the N- and C-terminal domains from hEC-SOD fused to hCuZnSOD, called FusNCZ and PseudoEC-SOD, respectively. We show that these proteins can be produced in large quantities in Escherichia coli, that they can be purified with high yields and that the characteristics of PseudoEC-SOD closely resemble those of hEC-SOD. Further, we extended our studies of the nature of the subunit interaction by investigating the involvement of the central domain.  (+info)

Both transcriptional and posttranscriptional mechanisms regulate human telomerase template RNA levels. (6/422)

The human telomerase RNA component (hTR) is present in normal somatic cells at lower levels than in cancer-derived cell lines. To understand the mechanisms regulating hTR levels in different cell types, we have compared the steady-state hTR levels in three groups of cells: (i) normal telomerase-negative human diploid cells; (ii) normal cells transfected with the human telomerase catalytic subunit, hTERT; and (iii) cells immortalized in vitro and cancer cells expressing their own endogenous hTERT. To account for the differences in steady-state hTR levels observed in these cell types, we compared the transcription rate and half-life of hTR in a subset of these cells. The half-life of hTR in telomerase-negative cells is about 5 days and is increased 1.6-fold in the presence of hTERT. The transcription rate of hTR is essentially unchanged in cells expressing exogenous hTERT, and the increased steady-state hTR level appears to be due to the increased half-life. However, the transcription rate of hTR is greatly increased in cells expressing endogenous hTERT, suggesting some overlap in transcriptional regulatory control. We conclude that the higher hTR level in cells expressing an endogenous telomerase can be a result of both increased transcription and a longer half-life and that the longer half-life might be partially a result of protection or stabilization by the telomerase catalytic subunit. The 4-week half-life of hTR in H1299 tumor cells is the longest half-life yet reported for any RNA.  (+info)

Occurrence of P-flavin binding protein in Vibrio fischeri and properties of the protein. (7/422)

In previous studies involving Photobacterium species we proposed that (i) P-flavin is the product of luciferase, (ii) the physiological function of the lux operon is not to produce light but to produce FP(390) (luxF protein), including its prosthetic group, P-flavin, and (iii) FP(390) reactivates oxidatively inactivated cobalamin-dependent methionine synthase similar to flavodoxin but at relatively high ionic strength. It seems difficult to extend this idea to all luminous bacteria because the luxF gene is not present in the lux operon in Vibrio or Xenorhabdus. But we predicted that a luciferase fragment which binds P-flavin should function like FP(390) in these species. In this study, we isolated P-flavin binding protein from Vibrio fischeri ATCC 7744. The obtained protein was a modified luciferase as expected, in which the beta-subunit was intact but about 25 amino acid residues at the C-terminus of the alpha-subunit were deleted and the prosthetic group was the fully reduced P-flavin. These results strongly support that the physiological function of the lux operon is as described above even in luminous bacteria other than Photobacterium species. We propose that chromophore B reported by Tu and Hastings [Tu, S.-C. and Hastings, J.W. (1975) Biochemistry 14, 1975-1980] is the reduced P-flavin.  (+info)

Bovine liver phosphoamidase as a protein histidine/lysine phosphatase. (8/422)

A 13-kDa phosphoamidase was isolated as a single band on SDS-PAGE from bovine liver. Its Stokes' radius, sedimentation coefficient, molecular mass, and optimal pH were estimated to be 1.6 nm, 1.8 s, 13 kDa, and 6.5, respectively. The enzyme released P(i) from 3-phosphohistidine, 6-phospholysine, and amidophosphate at rates of 0.9, 0.6, and 2.6 micromol/min/mg protein, respectively. However, it did not dephosphorylate phosphocreatine, N(omega)-phosphoarginine, imidodiphosphate, or O-phosphorylated compounds including inorganic pyrophosphate. It also dephosphorylated succinic thiokinase and nucleoside diphosphate kinase autophosphorylated at His residues, indicating that it works as a protein histidine phosphatase. A thiol reagent, 30 microM N-ethylmaleimide, depressed the activity by half, while a thiol compound, 2-mercaptoethanol, protected the enzyme from heat-inactivation. Five millimolar divalent cations, such as Mg2+ and Mn2+, and 5 mM EDTA, had no effect on the activity.  (+info)

Complete information for PSAP gene (Protein Coding), Prosaposin, including: function, proteins, disorders, pathways, orthologs, and expression. GeneCards - The Human Gene Compendium
Recombinant Human TEC, catalytic domain [359-631 amino acids] was expressed as N-terminal GST-fusion protein (59 kDa) usingbaculovirusexpression system. GST-TEC was purified by using glutathione sepharose chromatography and anion exchange chromatography.
A.T. Hanke, M.E. Klijn, P.D.E.M. Verhaert, L.A.M. van der Wielen, M. Ottens, M.H.M. Eppink, E.J.A.X. van de Sandt, Prediction of protein retention times in hydrophobic interaction chromatography by robust statistical characterization of their atomic-level surface properties, Biotechnol Prog. (2015) 1520-6033, http://dx.doi.org/10.1002/btpr.2219 ...
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Hydrophobic Interaction Chromatography is a separation technique that uses the properties of hydrophobicity to separate proteins from one another. In this type of chromatography, hydrophobic groups such as phenyl, octyl, or butyl, are attached to the stationary column. Proteins that pass through the column that have hydrophobic amino acid side chains on their surfaces are able to interact with and bind to the hydrophobic groups on the column. HIC separations are often designed using the opposite conditions of those used in ion exchange chromatography. In this separation, a buffer with a high ionic strength, usually ammonium sulfate, is initially applied to the column. The salt in the buffer reduces the solvation of sample solutes thus as solvation decreases, hydrophobic regions that become exposed are adsorbed by the medium. ...
Agarose偶联6X His tag®抗体(ab1231)经IP实验严格验证,实验条件参看说明书。Abcam对所有产品均提供质保服务和专属技术支持,中国75%以上现货。
Its only fair to share… Sacubitril, AHU 377 NEPRILYSIN INHIBITOR FOR HEART FAILURE CAS 149709-62-6 CAS SODIUM SALT 149690-05-1 (2R,4S)-5-(biphenyl-4-yl)-4-((3-carboxypropionyl)amino)-2-methylpentanoic acid ethyl ester 5-(Biphenyl-4-yl)-4(S)-(3-carboxypropionamido)-2(R)-methylbutyric acid ethyl ester N-(3-carboxy-1-oxopropyl)-(4S)-(p-phenylphenylmethyl)-4-amino-2R-methyl butanoic acid ethyl ester [1,1′-Biphenyl]-4-pentanoic acid, γ-[(3-carboxy-1-oxopropyl)amino]-α-methyl-, α-ethyl ester, (αR,γS)- [1,1′-Biphenyl]-4-pentanoic acid, γ-[(3-carboxy-1-oxopropyl)amino]-α-methyl-, ethyl ester, [S-(R*,S*)]- (2R,4S)-4-[(3-Carboxy-1-oxopropyl)amino]-4-[(p-phenylphenyl)methyl]-2-methylbutanoic acid ethyl ester (2R,4S)-5-(Biphenyl-4-yl)-4-[(3-carboxypropionyl)amino]-2-methylpentanoic acid ethyl ester Formula C24H29NO5 MW 411.49 …. Read more ...
This was a randomized, double-blind, placebo-controlled, dose escalation study to assess the safety and tolerability of 100 mg and 200 mg of inhaled Alpha-1 HC administered once a day for three weeks in subjects aged 18 years and older with cystic fibrosis (CF). The treatment duration in this study was intended to provide multi-dose safety information prior to proceeding to longer durations of exposure ...
This webcast presents a series experiments in which in-process mAb material was spiked with MVM-MVP and processed through Anion Exchange and Cation Exchange, and a a viral clearance study was performed on Hydrophobic Interaction Chromatography (HIC) resins
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In 1973, Roosevelt Cole, on his own behalf and on behalf of all present and future black inmates in the Panola County jail, sued the individual members of the Board of Supervisors of Panola County, Mississippi and the Sheriff for neglecting to look into the state of the prisons as required by Mississippi statute. The plaintiff complained of the unsanitary and unsafe conditions, the absence of fair procedures, physical abuse, denial of religious and political freedoms, inadequate medical attention, denial of a right to fair trial, lack of educational, recreational, and rehabilitation facilities, denial of essential communication due to poverty, inadequate diet, and racial discrimination against black inmates ...
Explore our array of standards for IEX, SEC (gel filtration), hydrophobic interaction chromatography or qualitative organic acid/carbohydrate analysis.
DUBLIN, Dec. 7, 2017 /PRNewswire/ --The Zeolite Molecular Sieve Market: Global Industry Analysis, Trends, Market Size and Forecasts up to 2023 report has ...
Windows 8 currently still in its pre-beta phase of development, has an official build number for 8176. Microsoft is slated to release the Beta of its next version of operating system sometime in late February. Today, a
Chromatography is a critical operational step in many pharmaceutical downstream processes. In this course, you will learn chromatography fundamentals, design, operations, key mechanisms, and performance testing. Laboratory exercises will provide hands-on experience in both Ion-exchange and hydrophobic interactions chromatography including equipment setup, column conditioning, performance testing and column operation. In addition, use of automation software and column packing will be demonstrated.
The present invention relates to functionalized molecular sieves that are useful as shape-selective adsorbents and catalysts. The inventive molecular sieves have a crystalline framework and include micropores of substantially uniform size and shape formed therein. These micropores contain one or more accessible organic moieties that are linked to the crystalline framework by a carbon-silicon ond. Shape selectivity of the inventive molecular sieves may be modulated by varying the size and shape of the micropores (which is a function of the particular molecular sieve being synthesized) and the organic moiety incoporated therein. In preferred embodiments, the molecular sieves of the present invention include crosslinked monomers of the formula wherein: X is a subsituted or unsubstituted moiety and R1 and R2 are each independently either oxygen, or a substituted or unsubstituted moiey, wherein the moiety is selected from the group consisting of C1-C20 alkyl, C2-C10 alkenyl, C2-C20 alkynyl, cycloalkyl,
The immunogen is Aflatoxin B1 (AFB1)-BSA conjugates. The antibody was affinity purified with an AFB1-Agarose column and competitively eluted by free AFB1. The antibody was conjugated to peroxidase (HRP) by reductive amination. This anti-AFB1 HRP conjugates could be utilized for detection and quantization of the food-borne mycotoxin AFB1. ...
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Glycoprotein Ib-IX-V (GPIb-IX-V) is a platelet adhesion receptor complex that initiates platelet aggregation. Glycoprotein Iba (GPIba) is the central component of the GPIb-IX-V complex, anchoring the complex to the cytoskeleton and harboring the binding site for von Willebrand factor (vWF). Previous studies suggest that the coagulation function in pigs differs from that in humans, especially with respect to the interaction between vWF and platelets. However, we have little knowledge about the function of porcine platelets, which is important with regard to studies of cardiovascular disease, clotting, and surgery that use pigs as animal models. To extend this information, we cloned and analyzed the porcine GPIba sequence. Porcine GPIba contains 1891 nucleotides and includes an open reading frame that encodes 627 amino acids. The nucleotide sequence showed 67% identity with human GPIba, whereas the deduced amino acid sequences were 59% identical. The vWF binding domain shares the highest identity ...
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GE Healthcare Sepharose Agarose Gel Filtration Media Sepharose CL-2B gel; 1000mL GE Healthcare Sepharose Agarose Gel Filtration Media Gel Filtration Media...
A method of preparing high purity procoagulant protein comprising the steps of (a) adsorbing a VIII:C/VIII:RP complex from a plasma or commercial concentrate source of factor VIII onto agarose beads bound to a monoclonal antibody specific to VIII:RP, (b) eluting VIII:C with a salt solution, (c) adsorbing the eluted VIII:C on an animohexyl agarose column and eluting the VIII:C with a salt solution.
Zeolite molecular sieve 3A manufacturer in China mainland,US $ 500 - 1,000 / Metric Ton, Chemical Auxiliary Agent, 88663-79-3, molecular sieves.Source from Zibo Yinghe Chemical Co., Ltd. on Alibaba.com.
... ,UltraPure™ Agarose resolves DNA and RNA fragments from 100 bp to >30 kb. UltraPure™ Agarose is a standard melting temperature multi-purpose agarose that is ideal for routine separation analysis. Its high gel strength specification ensures that gels will not break during handling nor d,medicine,medical supply,medical supplies,medical product
Agarose偶联VSV-G tag抗体(ab3843)经WB, ELISA, ICC实验严格验证,被1篇文献引用,实验条件参看说明书。Abcam对所有产品均提供质保服务和专属技术支持,中国75%以上现货。
"Separation of Newly-Synthesized RNA by Organomercurial Agarose Affinity Chromatography". J. Biochem. 81 (5): 1247-1252. PMID ... As a result, organomercurial agarose gel or gel beads are used to isolate thiolated compounds (such as thiouridine) in a ...
"Characterization of proteins and other macromolecules by agarose gel chromatography". Journal of Chromatography A. 152 (1): 21- ...
"Purification of a sialyltransferase from bovine colostrum by affinity chromatography on CDP-agarose". J. Biol. Chem. 252 (7): ...
... by affinity chromatography on agarose-linked egg-yolk lipoprotein". Biochim. Biophys. Acta. 351: 155-171. doi:10.1016/0005-2795 ...
... of a cortical complex containing two unconventional actins from Acanthamoeba by affinity chromatography on profilin-agarose". ... The Arp2/3 complex was named after it was identified in 1994 by affinity chromatography from Acanthamoeba castellanii, though ... a complex network of interacting proteins detected by F-actin affinity chromatography". The Journal of Cell Biology. 109 (6 Pt ...
... is often used to immobilize proteins by coupling them to reagents such as agarose for affinity chromatography ... These groups are reacted with primary amines in order to couple the protein onto the agarose matrix, as shown in the figure. ... Cyanogen bromide is also often used because it reacts with the hydroxyl groups on agarose to form cyanate esters and ... Also, cyanogen bromide activation involves the attachment of a ligand to agarose by an isourea bond, which is positively ...
The ability of lactoferrin to bind DNA is used for its isolation and purification using affinity chromatography with columns ... containing immobilized DNA-containing sorbents, such as agarose with the immobilized single-stranded DNA. Lactoferrin's primary ...
The agarose was chosen as the gel matrix because it has large pores allowing free passage and separation of proteins, but ... Some variants of affinity immunoelectrophoresis are similar to affinity chromatography by use of immobilized ligands. The open ... Agarose as 1% gel slabs of about 1 mm thickness buffered at high pH (around 8.6) is traditionally preferred for the ... Immunoprecipitates may be seen in the wet agarose gel, but are stained with protein stains like Coomassie Brilliant Blue in the ...
In: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences. Band 820, Nummer 1, Juni 2005, ... It is used in SDS-PAGE, agarose gel electrophoresis and histologic staining, e.g. staining of growth lines in bones. M. R. ... N. Volpi, F. Maccari: Detection of submicrogram quantities of glycosaminoglycans on agarose gels by sequential staining with ... Simultaneous detection of submicrogram quantities of hyaluronic acid and dermatan sulfate on agarose-gel by sequential staining ...
The stationary phase is a resin composed of beads, usually of cross-linked agarose, packed into a cylindrical glass or plastic ... Thin layer chromatography (TLC)[edit]. Further information: Thin layer chromatography. Thin layer chromatography (TLC) is a ... Gas chromatography[edit]. Further information: Gas chromatography. Gas chromatography (GC), also sometimes known as gas-liquid ... Affinity chromatography[edit]. Further information: Affinity chromatography. Affinity chromatography[14] is based on selective ...
... /agarose, combined with some form of activation chemistry, is also used to immobilize enzymes, antibodies and other ... Gel permeation chromatography Ahmed, Hafiz (2004). Principles and Reactions of Protein Extraction, Purification, and ... Sepharose is a tradename for a crosslinked, beaded-form of agarose, a polysaccharide polymer material extracted from seaweed. ... Its brand name is derived from Separation-Pharmacia-Agarose. A common application for the material is in chromatographic ...
Some common carbohydrate molecules that is used in lectin affinity chromatography are Con A-Sepharose and WGA-agarose.[22] ... Weak affinity chromatography[edit]. Weak affinity chromatography[28] (WAC) is an affinity chromatography technique for affinity ... A chromatography column containing nickel-agarose beads used for purification of proteins with histidine tags ... Boronate affinity chromatography[edit]. Boronate affinity chromatography consists of using boronic acid or boronates to elute ...
... making it more suitable for analytical techniques such as agarose gel electrophoresis, and chromatography. It is used in ... In gel electrophoresis, a sample of DNA is first "loaded" onto a slab of agarose gel (literally pipetted into small wells at ... Agarose gel electrophoresis DNA sequencing Genetic fingerprinting PCR Restriction fragment length polymorphism Hartl, Daniel L ... After restriction digest, DNA can then be analysed using agarose gel electrophoresis. ...
Another method of separation uses a sequence of freezing, thawing, and compression of SWNTs embedded in agarose gel. This ... Moreover, SWNTs can be separated by the column chromatography method. Yield is 95% in semiconductor type SWNT and 90% in ...
Agarose gel based medium contain large pores as well but their substitution ability is lower in comparison to dextrans. The ... Micellar liquid chromatography. Ion chromatography (or ion-exchange chromatography) is a chromatography process that separates ... High performance liquid chromatography. Aqueous Normal Phase Chromatography. Size exclusion chromatography. ... Anion exchange chromatography retains anions using positively charged functional group: R-X. +. A. −. +. M. +. B. −. ⇄. R-X. + ...
Using a low voltage (~10 V/cm) to minimize the risk for heat damage, electricity is run across an agarose gel. This technique ... Some of the methods are similar to affinity chromatography by use of immobilized ligands. Currently, there is ongoing research ... This method differs from the traditional agarose gel electrophoresis by utilizing a higher voltage to facilitate a shorter run ... A type of electrophoretic mobility shift assay (AMSA), agarose gel electrophoresis is used to separate protein-bound amino acid ...
... acid Agarose gel electrophoresis Food rheology Gel electrophoresis Gel filtration chromatography Gel permeation chromatography ... Natural hydrogel materials are being investigated for tissue engineering; these materials include agarose, methylcellulose, ...
... dextran sulfate-agarose chromatography, gel filtration on ACA-44, and DEAE-Sephacel chromatography. The structure (primary ...
Cellulose, dextran, agarose and other insoluble complexes are unaffected because they compose inert matrices, hence why they ... To ensure that the resin is protonated and positively charge, the chromatography should be performed at least 2 pH units below ... DEAE-Sepharose, DEAE-650 and DEAE-Sephadex are commonly used in chromatography. DEAE-C is a weak anion exchanger. This exchange ... Size-exclusion chromatography Stationary phase "Whatman® anion exchange cellulose DE52". Sigma-Aldrich. Retrieved 11 May 2016. ...
... dextran covalently linked to agarose). For affinity chromatography, beaded agarose is the most commonly used matrix resin for ... and low EEO agarose is therefore generally preferred for use in agarose gel electrophoresis of nucleic acids. Zero EEO agaroses ... Agarose is a useful material for chromatography because it does not absorb biomolecules to any significant extent, has good ... Examples of agarose-based matrix for gel filtration chromatography are Sepharose and WorkBeads 40 SEC (cross-linked beaded ...
Agarose gel based medium contain large pores as well but their substitution ability is lower in comparison to dextrans. The ... Ion chromatography (or ion-exchange chromatography) is a chromatography process that separates ions and polar molecules based ... This type of chromatography is further subdivided into cation exchange chromatography and anion-exchange chromatography. ... or in chromatography columns. Thin layer chromatography or column chromatography share similarities in that they both act ...
... coli β-galactosidase is accomplished by affinity chromatography using p-aminobenyl-1-thio-β-D-galactopyranosyl agarose as the ... Lectin affinity chromatography is a form of affinity chromatography where lectins are used to separate components within the ... Weak affinity chromatography (WAC) is a affinity chromatography technique for affinity screening in drug development. WAC is an ... Affinity chromatography is an excellent choice for the first step in purifying a protein or nucleic acid from a crude mixture. ...
Continuous chromatography, more precisely periodic counter-current chromatography, enormously increases the productivity of the ... It is also widely utilized coupled to magnetic, latex and agarose beads. Protein A is often immobilized onto a solid support ... Affinity Chromatography (PDF). Vol. 1: Antibodies (AF ed.). GE Healthcare. 2016. p. 48. "A Pathogen's Swiss Army Knife". Small ... Albeit the long history of protein A chromatography for the production of antibodies, the process is still being improved today ...
... other gels with size fractionation properties include agarose and polyacrylamide. A short review of these developments has ... Size-exclusion chromatography (SEC), also known as molecular sieve chromatography, is a chromatographic method in which ... Modern Size Exclusion Chromatography, Practice of Gel Permeation and Gel Filtration Chromatography, 2nd ed.; Wiley: NY, 2009. ... the technique is known as gel-filtration chromatography, versus the name gel permeation chromatography, which is used when an ...
Commercial gels like PLgel, Sephadex, Bio-Gel (cross-linked polyacrylamide), agarose gel and Styragel are often used based on ... Gel permeation chromatography (GPC) is a type of size exclusion chromatography (SEC), that separates analytes on the basis of ... Gel permeation chromatography is conducted almost exclusively in chromatography columns. The experimental design is not much ... Gel permeation chromatography (GPC) has become the most widely used technique for analyzing polymer samples in order to ...
Journal of Liquid Chromatography & Related Technologies. 20 (16-17): 2857-2872. doi:10.1080/10826079708005597.. ... Agarose gel electrophoresis. *Capillary electrochromatography. *Capillary electrophoresis. *Dielectrophoresis. *Difference gel ...
I have a quick question regarding agarose bead size in affinity chromatography, specifically nickel-coated beads for ... Agarose bead size in affinity chromatography - posted in Protein Expression and Purification: Hello everybody, ... RNA agarose gel blurry fuzzy. RNase contaminaton? Started by qpwoei4756, 16 Nov 2015 rna, agarose, contamination * 2 replies ... Chromatography machines? Started by wincel, 27 Jan 2016 chromatography, virus and 1 more... * 2 replies ...
... He,, X. ... can be separated from other isoflavonoids by adsorption chromatography on the cross-linked 12% agarose gel Superose 12 ... Isoflavonoid separation, Agarose cross-linker, Puerarin, Acetic acid quenching Identifiers. URN: urn:nbn:se:uu:diva-70502OAI: ... Thus, no useful separation can be achieved with non-cross-linked 12% agarose gel media. Symmetric elution profiles at high ...
Suitable for affinity chromatography of various cAMP-respon ... hydroxy group immobilized on agarose by an aminohexylamino ... Suitable for affinity chromatography of various cAMP-responsive proteins, especially those which tolerate modification of the ... 8-AHA-2-O-Me-cAMP-Agarose 8- (6- Aminohexylamino)- 2- O- methyladenosine- 3, 5- cyclic monophosphate; immobilized on ... The second messenger cyclic AMP with a methylated ribose 2-hydroxy group immobilized on agarose by an aminohexylamino spacer ...
Affinity chromatography on CaM-agarose. For interaction of AtDEG15 from inclusion bodies with calmodulin, 70 μl CaM-agarose ... S1), affinity chromatography on CaM-agarose (Fig. 2 and Fig. S2), bioinformatic analyses (Fig. 3) and CaM-AtDEG15 peptide ... S3). When the same purified protein fraction was analysed by affinity chromatography on CaM-agarose (Fig. 2A), the full length ... We repeated the affinity chromatography on CaM-agarose using recombinant AtDEG15 expressed in E. coli under the control of an ...
Agarose Bead Technologies ABT manufactures agarose resins for separation purification of biomolecules. Size Exclusion, Ion ...
Biotin Agarose Resin is used for purification or removal of avidin or streptavidin samples. Biotin is immobilized through a ... Biotin Agarose Resin is used for purification or removal of avidin or streptavidin samples. Biotin is immobilized through a ...
Chromatography on mannose agarose indicating an in vivo interaction of both proteins. The same method applied to a ... Chromatography on mannose agarose indicating an in vivo interaction of both. Chromatography on mannose agarose indicating an in ... The positive control additionally contained purified LecB protein (concentr.Chromatography on mannose agarose indicating an in ... Moreover, OprF could be isolated from the outer membrane fraction by His-tagged LecB immobilized on Ni-NTA agarose and could ...
Chelating-agarose chromatography. The solution then was applied to a Ni2+-iminodiacetic-agarose column (0.5 × 4.0 cm; Sigma) ... Hydrophobic chromatography or both Mono Q and hydrophobic chromatographies were repeated once if more than one band appeared. ... Phenyl-Superose chromatography. Active fractions recovered from the Mono Q column chromatography were pooled, diluted 10-fold ... Mono Q chromatography. The dialyzed solution was filtered through a Millipore 0.22-μm filter and applied to a Mono Q HR 5/5 ...
Ni2+ agarose chromatography. Fifteen milligrams of extract in lysis buffer plus 5 mM imidazole was bound to 100 μL of Ni2+‐NTA ... and Rad9 was purified by affinity chromatography on Ni2+ agarose (Figure 7). This protocol reproducibly resulted in most of ... Using Ni2+ affinity chromatography, and antibodies specific to Rad24, Rad17, Mec3 and Rad53 we have demonstrated that after DNA ... Native yeast protein extracts were prepared for chromatography as follows: cell pellets were washed with de‐ionised sterile ...
... resolution agarose based chromatography resins for protein purification Purolite Life Sciences introduces a new line of agarose ... IonQuest Ion Chromatography One single IonQuest system will accommodate your requirements for now and the future. There are no ... Waters and DAISO sign marketing agreement for bulk process chromatography media Waters Corporation and DAISO CO., LTD. jointly ... High-Performance Praesto® Protein A and Ion Exchange Chromatography Resins paired with Leading OPUS® Technology Purolite Life ...
Cuatrecasas P (1970) Protein purification by affinity chromatography. Derivativations of agarose and polyacrylamide beads. J ...
4. Chromatography of Fractions from Green-19 Agarose. The diluted sample containing synthase activity is loaded onto the ACP- ... C. Reactive Green 19 Agarose Chromatography. Fraction D is diluted with "20 buffer" to a final volume of 1650 ml. Fraction D is ... The agarose is packed at 200 ml/hr, until the last of the "50 buffer" reaches the top of the Green 19-agarose bed. The synthase ... The Green 19-agarose is filtered on a sintered glass funnel and washed with 200 ml of "50 buffer". The washed Green 19-agarose ...
d) Keparin agarose chromatography. The clarified sample was applied to a column of heparin agarose (15 1.6 cm) previously ... The yield of partially pure APP from the 300 mM heparin agarose eluent was 5.5 μg (Bradford assay) per gram of wet CHO cell ... High pressure liquid chromatography (HPLC) is particularly useful in this regard. In applying this technique, a fluorescently ... c) Purification of solubilized holo-APP 69S by strong anion exchange chromatography. The above supernatant containing holo-APP ...
Minibodies and Multimodal Chromatography Methods. by Pete Gagnon, Mark A. Sherman, Chia-Wei Cheung, Eric J. Lepin, Anna M. Wu, ... Mixed-mode chromatography sorbents can save time and money by reducing the number of steps required to purify recombinant ... Protein A affinity chromatography is traditionally used as the capture step for monoclonal antibodies (MAbs) (1,2,3). It yields ... Affinity chromatography is one of the simplest and most effective methods for purifying protein and peptide therapeutics, ...
chromatography; atomic force microscopy; adsorbents; particle technology; agarose hydrogel. Academic Unit/School:. Faculty of ... Manufacturing of agarose-based chromatographic adsorbents with controlled pore and particle size.. In: 69th Annual Technical ... Solutions of agarose containing different amounts of NaCl were emulsified at elevated temperature in mineral oil using a high- ... Manufacturing of agarose-based chromatographic adsorbents with controlled pore and particle size ...
The inactivated, chromatography-purified and concentrated BTV antigens are made into vaccine by dilution in a buffer solution ... The comb is inserted and the agarose is allowed to solidify on a level surface for 30-60 minutes. The comb and the tape are ... NuSieve 3/1 agarose: FMC Bioproducts, Rockland, Maine, USA. SYBR® Safe DNA gel stain: Available from Invitrogen (Cat No. S33102 ... Pour the tank buffer into the electrophoresis apparatus and insert the tray with the agarose so that the buffer covers the ...
Cuatrecasas, P., and Parikh, I., 1972, Adsorbents for affinity chromatography. Use of Nhydroxysuccinimide esters of agarose, ...
... purified enzyme after Ni-nitrilotriacetic acid agarose chromatography. Native PAGE of the purified enzyme. Lanes: 5, purified ... 24). Chromosomal DNA extracted from E. coli W3110 was partially digested with Sau3AI and size-fractionated (2-4 kb) by agarose ... Agarose (ultra PURE, GIBCO/BRL) was used for preparation of M9 plates. Well-separated colonies on the M9 plates were in ...
Covalent coupling of nucleotides to agarose for affinity chromatography. Biochim. Biophys. Acta, 304: 231-235.PubMedCrossRef ...
Glycobiology, Affinity Chromatography. Matrix Conjugate. Lectins. Sugar Specificity. Mannose, Glucose. Conjugate. Agarose. ... Agarose bound* Banana Lectin is prepared using our affinity-purified lectins. Heat stable, cross-linked 4% agarose beads with a ... Our agarose bound lectins are supplied at a constant concentration of lectin per ml of settled beads. The concentration for ... The technique we have developed to couple lectins to agarose beads inserts a hydrophilic spacer arm between the lectin and the ...
Glycobiology, Affinity Chromatography. Matrix Conjugate. Lectins. Sugar Specificity. [GlcNAc]1-3, N-Acetylglucosamine. ... Agarose bound* tomato lectin is prepared using our affinity-purified lectins. Heat stable, cross-linked 4% agarose beads with a ... Our agarose bound lectins are supplied at a constant concentration of lectin per ml of settled beads. The concentration for ... The technique we have developed to couple lectins to agarose beads inserts a hydrophilic spacer arm between the lectin and the ...
Agarose Matrices. Vector Laboratories affinity binding matrices are made using 4% agarose gel beads that have been crosslinked ...
Some common carbohydrate molecules that is used in lectin affinity chromatography are Con A-Sepharose and WGA-agarose.[22] ... Weak affinity chromatography[edit]. Weak affinity chromatography[28] (WAC) is an affinity chromatography technique for affinity ... A chromatography column containing nickel-agarose beads used for purification of proteins with histidine tags ... Boronate affinity chromatography[edit]. Boronate affinity chromatography consists of using boronic acid or boronates to elute ...
Purchase Affinity Chromatography and Biological Recognition - 1st Edition. Print Book & E-Book. ISBN 9780121665807, ... Mild Hydrophobic Chromatography. Protein Adsorption Hysteresis on Hydrophobic Agaroses. Effect of Matrix on Biospecific Complex ... Affinity Chromatography of Interferons-A Novel Application for Calmodulin-Sepharose. Dextran-Sepharose Affinity Chromatography ... Subunit Exchange Chromatography: Principles and Applications. Affinity Chromatography Studies of Cooperative Interactions in ...
PAGE and agarose gel electrophoresis. - Thin-layer chromatography. - HPLC. - Western and southern blots. - Identification of ...
  • Agarose is one of the two principal components of agar, and is purified from agar by removing agar's other component, agaropectin. (wikipedia.org)
  • Electroendosmosis is a reason agarose is used preferentially over agar as agaropeptin in agar contains a significant amount of negatively charged sulphate and carboxyl groups. (wikipedia.org)
  • This inhibitor with a hydrocarbon chain is commonly known as the spacer between the agarose bead and the target molecule. (wikipedia.org)
  • Agarose Bead Technology features Protein A and G, Ni NTA, Glutathione, Streptavidin and a customized media service. (botw.org)