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)Chromatography, High Pressure Liquid: Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.Chromatography, 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.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)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.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.Molecular Weight: The sum of the weight of all the atoms in a molecule.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.Electrophoresis, Polyacrylamide Gel: Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.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.Antibody Affinity: A measure of the binding strength between antibody and a simple hapten or antigen determinant. It depends on the closeness of stereochemical fit between antibody combining sites and antigen determinants, on the size of the area of contact between them, and on the distribution of charged and hydrophobic groups. It includes the concept of "avidity," which refers to the strength of the antigen-antibody bond after formation of reversible complexes.Binding Sites: The parts of a macromolecule that directly participate in its specific combination with another molecule.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)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.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.Mass Spectrometry: An analytical method used in determining the identity of a chemical based on its mass using mass analyzers/mass spectrometers.Substrate Specificity: A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.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.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)SepharoseRecombinant Proteins: Proteins prepared by recombinant DNA technology.Gas Chromatography-Mass Spectrometry: A microanalytical technique combining mass spectrometry and gas chromatography for the qualitative as well as quantitative determinations of compounds.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.Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.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.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.Ligands: 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)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).Peptide Fragments: Partial proteins formed by partial hydrolysis of complete proteins or generated through PROTEIN ENGINEERING techniques.Models, Molecular: Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.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.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.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.Protein 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).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)Cell Line: Established cell cultures that have the potential to propagate indefinitely.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.Macromolecular 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.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.Carrier Proteins: Transport proteins that carry specific substances in the blood or across cell membranes.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.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.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.Protein Structure, Tertiary: 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.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.Hydrolysis: The process of cleaving a chemical compound by the addition of a molecule of water.Cell Membrane: The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.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.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.Carbohydrate Sequence: The sequence of carbohydrates within POLYSACCHARIDES; GLYCOPROTEINS; and GLYCOLIPIDS.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).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.Liver: A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.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)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.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.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.Surface Plasmon Resonance: A biosensing technique in which biomolecules capable of binding to specific analytes or ligands are first immobilized on one side of a metallic film. Light is then focused on the opposite side of the film to excite the surface plasmons, that is, the oscillations of free electrons propagating along the film's surface. The refractive index of light reflecting off this surface is measured. When the immobilized biomolecules are bound by their ligands, an alteration in surface plasmons on the opposite side of the film is created which is directly proportional to the change in bound, or adsorbed, mass. Binding is measured by changes in the refractive index. The technique is used to study biomolecular interactions, such as antigen-antibody binding.Bacterial Proteins: Proteins found in any species of bacterium.Recombinant Fusion Proteins: 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.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).Radioligand Assay: Quantitative determination of receptor (binding) proteins in body fluids or tissue using radioactively labeled binding reagents (e.g., antibodies, intracellular receptors, plasma binders).Isoelectric Point: The pH in solutions of proteins and related compounds at which the dipolar ions are at a maximum.Mutagenesis, Site-Directed: Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion.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.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)Glycoproteins: Conjugated protein-carbohydrate compounds including mucins, mucoid, and amyloid glycoproteins.Antibodies, Monoclonal: Antibodies produced by a single clone of cells.Drug Stability: The chemical and physical integrity of a pharmaceutical product.Spectrometry, Fluorescence: Measurement of the intensity and quality of fluorescence.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)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)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)Methods: A series of steps taken in order to conduct research.Receptors, Cell Surface: Cell surface proteins that bind signalling molecules external to the cell with high affinity and convert this extracellular event into one or more intracellular signals that alter the behavior of the target cell (From Alberts, Molecular Biology of the Cell, 2nd ed, pp693-5). Cell surface receptors, unlike enzymes, do not chemically alter their ligands.Cricetinae: A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.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.PolysaccharidesSwine: 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).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)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.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.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.Dose-Response Relationship, Drug: The relationship between the dose of an administered drug and the response of the organism to the drug.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.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.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.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.CHO Cells: CELL LINE derived from the ovary of the Chinese hamster, Cricetulus griseus (CRICETULUS). The species is a favorite for cytogenetic studies because of its small chromosome number. The cell line has provided model systems for the study of genetic alterations in cultured mammalian cells.Calcium: A basic element found in nearly all organized tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes.Carbohydrate Conformation: The characteristic 3-dimensional shape of a carbohydrate.TritiumAdenosine Triphosphate: An adenine nucleotide containing three phosphate groups esterified to the sugar moiety. In addition to its crucial roles in metabolism adenosine triphosphate is a neurotransmitter.Hydroxyapatites: 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)Membrane Proteins: 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.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).Sequence Alignment: The arrangement of two or more amino acid or base sequences from an organism or organisms in such a way as to align areas of the sequences sharing common properties. The degree of relatedness or homology between the sequences is predicted computationally or statistically based on weights assigned to the elements aligned between the sequences. This in turn can serve as a potential indicator of the genetic relatedness between the organisms.Thermodynamics: A rigorously mathematical analysis of energy relationships (heat, work, temperature, and equilibrium). It describes systems whose states are determined by thermal parameters, such as temperature, in addition to mechanical and electromagnetic parameters. (From Hawley's Condensed Chemical Dictionary, 12th ed)Oligopeptides: Peptides composed of between two and twelve amino acids.Cross-Linking Reagents: 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.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.Peptide 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.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.Magnesium: A metallic element that has the atomic symbol Mg, atomic number 12, and atomic weight 24.31. It is important for the activity of many enzymes, especially those involved in OXIDATIVE PHOSPHORYLATION.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.Hot Temperature: Presence of warmth or heat or a temperature notably higher than an accustomed norm.Phosphorylation: The introduction of a phosphoryl group into a compound through the formation of an ester bond between the compound and a phosphorus moiety.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.Protein Structure, Secondary: The level of protein structure in which regular hydrogen-bond interactions within contiguous stretches of polypeptide chain give rise to alpha helices, beta strands (which align to form beta sheets) or other types of coils. This is the first folding level of protein conformation.Cytosol: Intracellular fluid from the cytoplasm after removal of ORGANELLES and other insoluble cytoplasmic components.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.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.Dimerization: The process by which two molecules of the same chemical composition form a condensation product or polymer.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).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)Erythrocytes: Red blood cells. Mature erythrocytes are non-nucleated, biconcave disks containing HEMOGLOBIN whose function is to transport OXYGEN.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.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.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)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.Brain: The part of CENTRAL NERVOUS SYSTEM that is contained within the skull (CRANIUM). Arising from the NEURAL TUBE, the embryonic brain is comprised of three major parts including PROSENCEPHALON (the forebrain); MESENCEPHALON (the midbrain); and RHOMBENCEPHALON (the hindbrain). The developed brain consists of CEREBRUM; CEREBELLUM; and other structures in the BRAIN STEM.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.Spectrophotometry: The art or process of comparing photometrically the relative intensities of the light in different parts of the spectrum.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.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.Plant Lectins: Protein or glycoprotein substances of plant origin that bind to sugar moieties in cell walls or membranes. Some carbohydrate-metabolizing proteins (ENZYMES) from PLANTS also bind to carbohydrates, however they are not considered lectins. Many plant lectins change the physiology of the membrane of BLOOD CELLS to cause agglutination, mitosis, or other biochemical changes. They may play a role in plant defense mechanisms.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.Glycoside HydrolasesCations, 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.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.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)Iodine Radioisotopes: Unstable isotopes of iodine that decay or disintegrate emitting radiation. I atoms with atomic weights 117-139, except I 127, are radioactive iodine isotopes.Crystallography, X-Ray: The study of crystal structure using X-RAY DIFFRACTION techniques. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Ammonium Sulfate: Sulfuric acid diammonium salt. It is used in CHEMICAL FRACTIONATION of proteins.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.Catalysis: The facilitation of a chemical reaction by material (catalyst) that is not consumed by the reaction.Epitopes: Sites on an antigen that interact with specific antibodies.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)DNA-Binding Proteins: Proteins which bind to DNA. The family includes proteins which bind to both double- and single-stranded DNA and also includes specific DNA binding proteins in serum which can be used as markers for malignant diseases.Acetonitriles: Compounds in which a methyl group is attached to the cyano moiety.Models, Chemical: Theoretical representations that simulate the behavior or activity of chemical processes or phenomena; includes the use of mathematical equations, computers, and other electronic equipment.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.Transfection: The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES.Histidine: An essential amino acid that is required for the production of HISTAMINE.Peptide Library: A collection of cloned peptides, or chemically synthesized peptides, frequently consisting of all possible combinations of amino acids making up an n-amino acid peptide.Isotope Labeling: 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.Blood Proteins: Proteins that are present in blood serum, including SERUM ALBUMIN; BLOOD COAGULATION FACTORS; and many other types of proteins.Enzyme Activation: Conversion of an inactive form of an enzyme to one possessing metabolic activity. It includes 1, activation by ions (activators); 2, activation by cofactors (coenzymes); and 3, conversion of an enzyme precursor (proenzyme or zymogen) to an active enzyme.Antibody Specificity: 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.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.Biological Transport: 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.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.Amino Acid Substitution: The naturally occurring or experimentally induced replacement of one or more AMINO ACIDS in a protein with another. If a functionally equivalent amino acid is substituted, the protein may retain wild-type activity. Substitution may also diminish, enhance, or eliminate protein function. Experimentally induced substitution is often used to study enzyme activities and binding site properties.Cysteine: A thiol-containing non-essential amino acid that is oxidized to form CYSTINE.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)Disaccharides: Oligosaccharides containing two monosaccharide units linked by a glycosidic bond.Chemistry, Physical: The study of CHEMICAL PHENOMENA and processes in terms of the underlying PHYSICAL PHENOMENA and processes.Physicochemical Phenomena: The physical phenomena describing the structure and properties of atoms and molecules, and their reaction and interaction processes.Cross Reactions: Serological reactions in which an antiserum against one antigen reacts with a non-identical but closely related antigen.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.DNA, Complementary: Single-stranded complementary DNA synthesized from an RNA template by the action of RNA-dependent DNA polymerase. cDNA (i.e., complementary DNA, not circular DNA, not C-DNA) is used in a variety of molecular cloning experiments as well as serving as a specific hybridization probe.Immunoglobulin G: The major immunoglobulin isotype class in normal human serum. There are several isotype subclasses of IgG, for example, IgG1, IgG2A, and IgG2B.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.Endopeptidases: A subclass of PEPTIDE HYDROLASES that catalyze the internal cleavage of PEPTIDES or PROTEINS.Anion Exchange Resins: High-molecular-weight insoluble polymers that contain functional cationic groups capable of undergoing exchange reactions with anions.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.Lysine: An essential amino acid. It is often added to animal feed.Chemistry Techniques, Analytical: Methodologies used for the isolation, identification, detection, and quantitation of chemical substances.Kidney: Body organ that filters blood for the secretion of URINE and that regulates ion concentrations.Mannose: A hexose or fermentable monosaccharide and isomer of glucose from manna, the ash Fraxinus ornus and related plants. (From Grant & Hackh's Chemical Dictionary, 5th ed & Random House Unabridged Dictionary, 2d ed)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.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.Saccharomyces cerevisiae: A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement.Proteomics: The systematic study of the complete complement of proteins (PROTEOME) of organisms.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.Glycosides: Any compound that contains a constituent sugar, in which the hydroxyl group attached to the first carbon is substituted by an alcoholic, phenolic, or other group. They are named specifically for the sugar contained, such as glucoside (glucose), pentoside (pentose), fructoside (fructose), etc. Upon hydrolysis, a sugar and nonsugar component (aglycone) are formed. (From Dorland, 28th ed; From Miall's Dictionary of Chemistry, 5th ed)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.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.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)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.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.Zinc: A metallic element of atomic number 30 and atomic weight 65.38. It is a necessary trace element in the diet, forming an essential part of many enzymes, and playing an important role in protein synthesis and in cell division. Zinc deficiency is associated with ANEMIA, short stature, HYPOGONADISM, impaired WOUND HEALING, and geophagia. It is known by the symbol Zn.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.Tumor Cells, Cultured: Cells grown in vitro from neoplastic tissue. If they can be established as a TUMOR CELL LINE, they can be propagated in cell culture indefinitely.HeLa Cells: The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for VIRUS CULTIVATION and antitumor drug screening assays.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)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.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.Protein Engineering: Procedures by which protein structure and function are changed or created in vitro by altering existing or synthesizing new structural genes that direct the synthesis of proteins with sought-after properties. Such procedures may include the design of MOLECULAR MODELS of proteins using COMPUTER GRAPHICS or other molecular modeling techniques; site-specific mutagenesis (MUTAGENESIS, SITE-SPECIFIC) of existing genes; and DIRECTED MOLECULAR EVOLUTION techniques to create new genes.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.Models, Biological: 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.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.Serum Albumin: A major protein in the BLOOD. It is important in maintaining the colloidal osmotic pressure and transporting large organic molecules.

Isolation and purification of rat mammary tumor peroxidase. (1/9384)

7,12-Dimethylbenz(a)anthracene-induced rat mammary tumors often contain high levels of the enzyme perioxidase, a putative marker of estrogen dependence. This enzyme can be effectively extracted with 0.5 M CaCl2, giving rise to a soluble peroxidase with a molecular weight of about 50,000 as determined by gel filtration. This is the same size as the estrogen-induced peroxidase of rat uterus but smaller than other mammalian peroxidases. Further purification of the rat mammary tumor peroxidase by concanavalin A-Sepharose chromatography and hydrophobic interaction chromatography on phenyl Sepharose provides a 640-fold purification of the enzyme.  (+info)

Involvement of poly (ADP-ribose)-polymerase in the Pax-6 gene regulation in neuroretina. (2/9384)

The quail Pax-6 gene is expressed from two promoters named P0 and P1. P0 promoter is under the control of a neuroretina-specific enhancer (EP). This enhancer activates the P0 promoter specifically in neuroretina cells and in a developmental stage-dependent manner. The EP enhancer binds efficiently, as revealed by southwestern experiments, to a 110 kDa protein present in neuroretina cells but not in Quail Embryos Cells and Retinal Pigmented Epithelium which do not express the P0-initiated mRNAs. To study the role of p110 in Pax-6 regulation, we have purified the p110 from neuroretina cells extracts. Based on the peptide sequence of the purified protein, we have identified the p110 as the poly(ADP-ribose) polymerase (PARP). Using bandshift experiments and footprinting studies, we present evidence that PARP is a component of protein complexes bound to the EP enhancer that increases the on rate of the protein complex formation to DNA. Using PARP inhibitors (3AB and 6.5 Hphe), we show that these products are able to inhibit EP enhancer activity in neuroretina cells. Finally, we demonstrate that these inhibitors are able to decrease the expression of the P0-initiated mRNA in the MC29-infected RPE cells which, in contrast to the RPE cells, accumulated the PARP in response to v-myc expression. Our results suggest that PARP is involved in the Pax-6 regulation.  (+info)

Comparison of the fibrin-binding activities in the N- and C-termini of fibronectin. (3/9384)

Fibronectin (Fn) binds to fibrin in clots by covalent and non-covalent interactions. The N- and C-termini of Fn each contain one non-covalent fibrin-binding site, which are composed of type 1 (F1) structural repeats. We have previously localized the N-terminal site to the fourth and fifth F1 repeats (4F1.5F1). In the current studies, using proteolytic and recombinant proteins representing both the N- and C-terminal fibrin-binding regions, we localized and characterized the C-terminal fibrin-binding site, compared the relative fibrin-binding activities of both sites and determined the contribution of each site to the fibrin-binding activity of intact Fn. By fibrin-affinity chromatography, a protein composed of the 10F1 repeat through to the C-terminus of Fn (10F1-COOH), expressed in COS-1 cells, and 10F1-12F1, produced in Saccharomyces cerevisiae, displayed fibrin-binding activity. However, since 10F1 and 10F1.11F1 were not active, the presence of 12F1 is required for fibrin binding. A proteolytic fragment of 14.4 kDa, beginning 14 residues N-terminal to 10F1, was isolated from the fibrin-affinity matrix. Radio-iodinated 14.4 kDa fibrin-binding peptide/protein (FBP) demonstrated a dose-dependent and saturable binding to fibrin-coated wells that was both competitively inhibited and reversed by unlabelled 14.4 kDa FBP. Comparison of the fibrin-binding affinities of proteolytic FBPs from the N-terminus (25.9 kDa FBP), the C-terminus (14.4 kDa) and intact Fn by ELISA yielded estimated Kd values of 216, 18 and 2.1 nM, respectively. The higher fibrin-binding affinity of the N-terminus was substantiated by the ability of both a recombinant 4F1.5F1 and a monoclonal antibody (mAb) to this site to maximally inhibit biotinylated Fn binding to fibrin by 80%, and by blocking the 90% inhibitory activity of a polyclonal anti-Fn, by absorption with the 25.9 kDa FBP. We propose that whereas the N-terminal site appears to contribute to most of the binding activity of native Fn to fibrin, the specific binding of the C-terminal site may strengthen this interaction.  (+info)

Purification and characterization of an alpha-galactosyltransferase from Trypanosoma brucei. (4/9384)

A membrane-associated galactosyltransferase from Trypanosoma brucei was purified 34000-fold by affinity chromatography on UDP-hexanolamine-Sepharosetrade mark. Using SDS/PAGE under reducing conditions, the isolated enzyme ran as a relatively broad band with apparent molecular masses of 53 kDa and 52 kDa, indicative of glycosylation and the existence of two isoforms. N-Glycosylation of the enzyme was subsequently confirmed using Western blotting and either specific binding of concanavalin A or peptide-N4-(N-acetylglucosaminyl)asparagine amidase digestion. The de-N-glycosylated enzyme ran with apparent molecular masses of 51 kDa and 50 kDa, indicative of a single N-glycosylation site. The galactosyltransferase exhibited a pH optimum at 7.2 and had a pronounced requirement for Mn2+ ions (KM=2.5 mM) for its action. The transferase activity was independent of the concentration of Triton X-100. The enzyme was capable of transferring galactose from UDP-galactose to a variety of galactose-based acceptors in alpha-glycosidic linkages. The apparent KM values for UDP-galactose and for the preferred acceptor substrate N-acetyl-lactosamine are 46 microM and 4.5 mM respectively. From these results we would like to suggest that the galactosyltransferase functions in the processing of terminal N-acetyl-lactosamine structures of trypanosomal glycoproteins.  (+info)

Enrichment of enzyme activity on deformylation of 1-NFK-lysozyme. (5/9384)

The formamide linkage of an inactive lysozyme derivative (1-NFK-lysozyme), formed by selective ozonization of tryptophan 62 in hen egg-white lysozyme [EC 3.2.1.17] was hydrolyzed with dilute acid faster in the frozen state at about --10 degrees than at 20 degrees. On hydrolysis of 1-NFK-lysozyme the low lytic activity increased to approximately 80% of that of native lysozyme. It is suggested that the binding ability associated with kynurenine 62 in the lysozyme derivative formed by this hydrolysis may be responsible for increase in enzymatic activity.  (+info)

Hydrophobic interaction of human, mouse, and rabbit interferons with immobilized hydrocarbons. (6/9384)

Interferons of human, mouse, and rabbit origin bind to straight chain hydrocarbons immobilized on agarose. The hydrophobic nature of binding is established by the following observations: (a) a positive correlation between the length of hydrocarbon ligand and the strength of interaction; (b) a stronger interaction with hydrocarbon ligands terminated with apolar rather than polar head groups; (c) a lack of dependence of binding on ionic strength and pH of the solvent; (d) a reversal of binding by ethylene glycol, a hydrophobic solute; (e) an increasing eluting efficacy of tetraalkylammonium ions with the length of their alkyl substituents. The hydrophobic interactions of human interferon underlie the efficiency of two-step chromatographic procedures. For example, human embryo kidney interferon can be purified about 3,600-fold by sequential chromatography on (a) concanavalin A-agarose, (b) octyl-agarose. Another two-step procedure: (a) concanavalin A-agarose, (b) L-tryptophan-agarose, gives about 10,000-fold purification. The overall recovery of interferon in both cases in close to 90%.  (+info)

Removal of non-specific serum inhibitors of haemagglutination of rubella virus by treatment with dodecylamine-gel. (7/9384)

The suitability of using dodecylamine-gel for removing the serum non-antibody-like inhibitors of haemagglutination by rubella was studied. Compared with kaolin and MnCl2/heparin treatment this new procedure appears to have a higher specificity since it removes the non-antibody-like inhibitors from serum without affecting the immunoglobulin level significantly. The potential application of this procedure in routine serological analysis for rubella virus infection is discussed.  (+info)

Purification of two dexamethasone-binding proteins from rat-liver cytosol. (8/9384)

Two dexamethasone-binding proteins have been purified from rat liver cytosol. The main purification steps are: precipitation by protamine sulphate, affinity chromatography on CH-Sepharose 4B to which 11-deoxycorticosterone is linked through a disulfide bond and DEAE-cellulose chromatography. Two binding components elute from the DEAE-cellulose column at 0.12 M and 0.2 M NaCl, respectively. By means of dodecylsulphate/polyacrylamide gel electrophoresis it was demonstrated that both components are composed predominantly of a single polypeptide with molecular weights of about 45 000 and 90 000. Antibodies to the two polypeptides have been elicited in rabbits. The antibodies to the 45 000-Mr polypeptide cross react with the 90 000-Mr component. Likewise the antibodies to the 90 000-Mr protein precipitate the 45 000-Mr polypeptide. Either of the two antibody preparations immunoprecipitates the major part (approximately 70%) of the dexamethasone-binding activity of the cytosol.  (+info)

*Affinity chromatography

Weak affinity chromatography (WAC) is a affinity chromatography technique for affinity screening in drug development. WAC is an ... Lectin affinity chromatography is a form of affinity chromatography where lectins are used to separate components within the ... Of many uses of affinity chromatography, one use of it is seen in affinity purification of albumin and macroglobulin ... Possibly the most common use of affinity chromatography is for the purification of recombinant proteins. Affinity ...

*Heparin

"Affinity Chromatography". Sigma-Aldrich. Archived from the original on 2016-05-07. "HiTrap Heparin HP". GE Healthcare Life ... Heparin has been used as a chromatography resin, acting as both an affinity ligand and an ion exchanger. Heparin's specific ... Segura MM, Kamen A, Garnier A (2008). "Purification of retrovirus particles using heparin affinity chromatography". Methods Mol ... "A novel purification strategy for tetrovirus gene therapy vectors using heparin affinity chromatography". Biotechnol Bioeng. 4 ...

*Protein A

Affinity Chromatography (PDF). Vol. 1: Antibodies (AF ed.). GE Healthcare. 2016. p. 48. "A Pathogen's Swiss Army Knife". Small ... Its isolation by affinity chromatography and its use as an immunosorbent for isolation of immunoglobulins". FEBS Letters. 28 (1 ... Continuous chromatography, more precisely periodic counter-current chromatography, enormously increases the productivity of the ... Protein A can bind with strong affinity to the Fc portion of immunoglobulin of certain species as shown in the below table. In ...

*KIAA0922

Purification of this endopeptidase by affinity chromatography". J. Biol. Chem. 251 (23): 7593-9. PMID 12173. "NCBI Gene: PREP ...

*TMTC4

All of these were detected through affinity chromatography. Ortholog space for TMTC4 spans a large portion of evolutionary time ...

*Type three secretion system

An additional approach for further purification uses affinity chromatography. Recombinant T3SS proteins that carry a protein ... the lysate is passed through a column coated with particles with high affinity to the tag (in the case of histidine tags: ...

*NanoCLAMP

Burgess RR, Watson JD (June 2017). "Gentle antibody-mimetic affinity chromatography with polyol-responsive nanoCLAMPs". Protein ... This property has been shown to be useful for purifying functional proteins and protein complexes by affinity purification. ... Thompson NE, Aronson DB, Burgess RR (1990). "Purification of eukaryotic RNA polymerase II by immunoaffinity chromatography. ... Thompson NE, Jensen DB, Lamberski JA, Burgess RR (2006). "Purification of protein complexes by immunoaffinity chromatography: ...

*Leukotriene C4 synthase

Söderström M, Morgenstern R, Hammarström S (1995). "Protein-protein interaction affinity chromatography of leukotriene C4 ...

*Microsomal glutathione S-transferase 1

Söderström M, Morgenstern R, Hammarström S (1995). "Protein-protein interaction affinity chromatography of leukotriene C4 ...

*Heterotetramer

Nickel affinity chromatography may also be employed for heterotetramer purification. Tetrameric protein "GO term: protein ... Ion-exchange chromatography is useful for isolating specific heterotetrameric protein assemblies, allowing purification of ...

*Downstream processing

Examples of operations include affinity, size exclusion, reversed phase chromatography, ion-exchange chromatography, ... Affinity chromatography often isolates and purifies in a single step. Fermentation (biochemistry) Separation process Unit ...

*Organomercury

"Separation of Newly-Synthesized RNA by Organomercurial Agarose Affinity Chromatography". J. Biochem. 81 (5): 1247-1252. PMID ...

*Microorganism

2009). "Production and Purification of Streptokinase by Protected Affinity Chromatography". Avicenna Journal of Medical ...

*Uridine monophosphate synthetase

Purification by tandem affinity chromatography of uridine-5'-monophosphate synthase". Biochemistry. 19 (20): 4699-706. doi: ... Union enthalpy and enthropy from the latter correspond to high-affinity ligands. Properties such as lipophilicity, solubility, ... Nonetheless, crystallographic analyses and the lack of S. cerevisae enzyme affinity to substrate analogues where the ...

*Annexin A2

Identification of two binding proteins obtained by Ca2(+)-dependent affinity chromatography". European Journal of Biochemistry ... Chung CY, Erickson HP (Jul 1994). "Cell surface annexin II is a high affinity receptor for the alternatively spliced segment of ...

*Maltose-binding protein

Protein X can then be separated from MBP by affinity chromatography. A first study of the relations between structure and ... develop the use of MBP as an affinity handle for the purification of foreign proteins and peptides by affinity chromatography ... In addition, MBP can itself be used as an affinity tag for purification of recombinant proteins. The fusion protein binds to ...

*Methylsterol monooxygenase

Kawata, S.; Trzaskos, J.M.; Gaylor, J.L. (1986). "Affinity chromatography of microsomal enzymes on immobilized detergent- ...

*1,3-alpha-L-fucosidase

"Purification of almond emulsin alpha-L-fucosidase I by affinity chromatography". Arch. Biochem. Biophys. 194 (2): 394-8. doi: ...

*Toluene dioxygenase

... purification of an iron-sulfur protein by affinity chromatography". Biochem. Biophys. Res. Commun. 91 (3): 1131-9. doi:10.1016/ ...

*FlAsH-EDT2

"Purification of Vicinal Dithiol-containing Proteins by Arsenical-Based Affinity Chromatography". In Sies, Helmut. Oxygen ... Such strong sulfur-arsenic bond can be, again, regulated by designing a peptide domain that exhibits higher affinity toward the ...

*S100A6

Identification of two binding proteins obtained by Ca2(+)-dependent affinity chromatography". Eur. J. Biochem. 195 (3): 795-800 ...

*Alpha-glucosidase

Sivikami, S.; Radhakrishnan, A.N. (1973). "Purification of rabbit intestinal glucoamylase by affinity chromatography on ... The substrate selectivity of alpha-glucosidase is due to subsite affinities of the enzyme's active site. Two proposed ...

*SMOX

Tsukada T, Furusako S, Maekawa S, Hibasami H, Nakashima K (1988). "Purification by affinity chromatography and characterization ...

*Tobacco etch virus

... followed by an affinity tag, such as a Polyhistidine-tag. Following affinity chromatography, the purified protein is then ... TEV protease cleaves at its recognition site, removing the affinity tag. This allows for affinity purification of proteins that ...

*Maltase

Sivikami, S.; Radhakrishnan, A.N. (1973). "Purification of rabbit intestinal glucoamylase by affinity chromatography on ...

*RBMX

... phosphoproteomic profiling of tyrosine phosphorylation sites from human T cells using immobilized metal affinity chromatography ...
[96 Pages Report] Check for Discount on United States Affinity Chromatography Columns Market Report 2017 report by QYResearch Group. In this report, the United States Affinity Chromatography Columns market...
In article ,CKBILy.7pG at ucdavis.edu, szsclark at hamlet.ucdavis.edu (Sonya Clark) writes: ,Dear Netters - I am purifying antibodies from a polyclonal prep by ,running them over a CNBr-activated Sepharose affinity column of my ,antigen. As CNBr-activated Sepharose is expensive &/or horrible to make, Well it isnt that expensive compared to the time saved by buying it (unless youre a graduate student whose time isnt valued much) and it isnt all that bad to make (N-methyl-pyrrolidone/Na2CO3 method ) if you have a fume hood but anyhow ,Id like to strip the column of the antigen currently bound, and re-use ,the sepharose to make another affinity column. the real point is that the activated bond from the CNBr is used up by the coupling procedure. Even if you could get the protein off, youre back to unactivated sepharose. Joe Mack mack at ncifcrf.gov Does anyone have a protocol ,to do this? Would eluting the column with harsh eluents (eg. ,Guanidine/urea/SDS) work? and would I then be able to ...
Our quantitative results differ considerably from those of Posewitz and Tempst (35), especially for the recovery of phosphopeptides from Fe3+-IDA with aqueous ammonia. We think that the difference is mainly due to the different aims in phosphopeptide capture MALDI compatibility (35) versus preparative-scale isolation in our study. We have eluted the peptides with a larger volume of dilute ammonia to achieve a sufficiently high pH in the column, and left the column for at least 5 min in these alkaline conditions (see "Materials and Methods"). Posewitz and Tempst note that with their eluting conditions, the eluate was "mildly alkaline," suggesting that their Fe3+-IMAC columns may not have reached the original pH of the eluant. They further note that "more than 10 volumes of the elution solvent, or significantly lower flow rate, was required to even begin desorbing the phosphorylated peptides." Our conditions are closer to this description. The recovery of 60-70% of the input is lower than ...
Affinity Chromatography (AC) is based on the specific adsorption of a molecule to a ligand or macromolecule. Most biomolecules can be purified on the basis of specific interaction between their chemical or biological structure and a suitable affinity ligand. Typical molecular pairs are enzymes and coenzymes or antigens and antibodies. AC packing materials have spacer ligands that are first attached to the substrate before a reversible adsorption of a specific biomolecule. The adsorbed molecule is then eluted through a competitive displacement or by a change in the conformation of the molecule through a change in pH or ionic strength. In contrast to other chromatographic methods, AC is highly selective and is mostly suitable for specific separation problems. Separation Methods Technologies silica-based Chemically Immobilized Biomolecules (CIB) columns are for high performance purification of other biomolecules such as proteins, enzymes and antibodies. In production of CIB columns, SMT utilizes its
GE Healthcare Epoxy-activated Sepharose 6B Medium Epoxy-activated Sepharose 6B Life Sciences:Protein Biology:Protein Extraction and Purification:Protein Purification:Affinity
Sugar moieties on the cell surface play one of the most important roles in cellular recognition. In order to elucidate the molecular mechanism of these cellular phenomena, assessment of the structure...
Nucleic acid affinity matrices that bear a large number of different nucleic acid affinity ligands allowing the simultaneous selection and removal of a large number of preselected nucleic acids from the sample. Methods of producing such affinity matrices are also provided. In general the methods involve the steps of a) providing a nucleic acid amplification template array comprising a surface to which are attached at least 50 oligonucleotides having different nucleic acid sequences, and wherein each different oligonucleotide is localized in a predetermined region of the surface, the density of the oligonucleotides is greater than about 60 different oligonucleotides per 1 cm2, and all of the different oligonucleotides have an identical terminal 3′ nucleic acid sequence and an identical terminal 5′ nucleic acid sequence. b) amplifying the multiplicity of oligonucleotides to provide a pool of amplified nucleic acids; and c) attaching the pool of nucleic acids to a solid support.
Generating your own affinity chromatography columns and resins allows for faster and cleaner protein purification. Pre-activated resins simplify the process.
In order to study individual biochemical compounds like proteins, DNA, or RNA, biochemists need to know how to purify these components from a complex mixture. This is especially important for biotechnology and pharmaceutical industries, which sell purified biochemicals as reagents or drugs to consumers. Do an experiment to purify DNA, RNA, or protein from a complex mixture (for purifying DNA, see the Science Buddies project Extracting Onion DNA). The source of the material can be a cell line, bacterial culture, plant extract, or yeast culture. Which purification strategies work best for your compound of interest? Can you use enzymes like protease, DNAse, or RNAse to test the product of your purification to see if it worked? Are there ways of altering the protocol to make it work better and increase the yield? For example, you could try changing detergent concentrations, salinity, or pH, or adding enzymes ...
Outcomes of comparative evaluations of enrichment methods for phosphopeptides depend highly on the experimental protocols used, the operator, the source of the affinity matrix, and the samples analyzed. Here, we attempt such a comparative study exploring a very large synthetic library containing thousands of serine, threonine, and tyrosine phosphorylated peptides, ... read more being present in roughly equal abundance, along with their nonphosphorylated counterparts, and use an optimized protocol for enrichment by TiO2 and Ti(4+)-immobilized metal affinity chromatography (IMAC) by a single operator. Surprisingly, our data reveal that there are minimal differences between enrichment of phosphopeptides by TiO2 and Ti(4+)-IMAC when considering biochemical and biophysical parameters such as peptide length, sequence surrounding the site, hydrophobicity, and nature of the amino acid phosphorylated. Similar results were obtained when evaluating a tryptic digest of a cellular lysate, representing a more ...
Creative BioMart, a world leading biotech company focused on supplying quality protein products including recombinant proteins, native proteins, GMP proteins, etc. and custom protein services, is pleased to enlarge its chromatography offerings to better serve scientists in the field of life sciences.. Chromatography is the collective term for a set of laboratory techniques for the separation of mixtures. The separation is based on differential partitioning between the mobile and stationary phases. Commonly used chromatography techniques include: gel filtration, ion exchange chromatography, hydrophobic interaction chromatography and affinity chromatography.. Upon this update, Creative BioMart now contains all types of chromatography at all scales of matrix including: cross-linked agarose, cross-linked cellulose, dextran, methacrylic and polystyrene. Customers can choose from affinity chromatography, antibody affinity chromatography to gel filtration chromatography and ion exchange chromatography, ...
An affinity matrix for use in affinity based molecular pull down and immunoprecipitation procedures. The affinity matrix comprises a polymeric support, a dye attached to a fraction of the polymeric support to enable optical detection of the polymeric support, and an affinity ligand other than the dye attached to a fraction of the polymeric support for the capture of a molecule. Also provided is a method for the isolation of a biomolecule from an aqueous solution. The method comprises combining the aqueous solution with an affinity matrix comprising a polymeric support and separating the affinity matrix from the aqueous solution. A dye is attached to a fraction of the polymeric support which enables optical detection and monitoring of the affinity matrix and, accordingly, reduces the likelihood of the loss of affinity matrix during the separation step. In addition, an affinity ligand other than the dye is also attached to a fraction of the polymeric support for the capture of the biomolecule.
Team:Freiburg_Bioware/Head}} {{:Team:Freiburg_Bioware/menu_home}} ,html> ,h1>Methods,/h1> ,/html> =Method Development= ==Purification of AAV particles== ===IMAC purification via Viral Brick: The Histidin Affinity Tag=== Protein tagging via Histidine Tags is a widely used method for protein purification: Multiple histidine residues (most commonly: Six) are being fused tot he end of the targeting protein. The high binding affinity of Histidine towards metal is being exploited for the purification of proteins via the so called „Immobilized Metal Ion Affinity Chromatography" (IMAC): Multiple histidine residues (most commonly: Six) are being fused to the end of the targeting protein. A cell extract containing the recombinant protein ist then applied to a collumn containing immobilized Ni2+-Ions. The His-tags covalently bind the Ni-Ions while other cellular proteins can be washed oft he collumn. The purified proteins can then be eluted with Imidazol, which displaces the histidine residues.(MC Smith ...
Affibody molecules are a class of small (ca.7 kDa) robust scaffold proteins with high potential as tracers for radionuclide molecular imaging in vivo. Incorporation of a cysteine-containing peptide-based chelator at the C terminus provides an opportunity for stable labelling with the radionuclide 99mTc. The use of a GGGC chelator at the C terminus has provided the lowest renal radioactivity retention of the previously investigated peptide-based chelators. Previously, it has also been demonstrated that replacement of the His6-tag with the negatively charged histidine-glutamate-histidine-glutamate-histidine-glutamate (HEHEHE)-tag permits purification of affibody molecules by immobilized metal ion affinity chromatography (IMAC) and provides low hepatic accumulation of radioactivity of conjugates site-specifically labelled at the C terminus using several different nuclides. We hypothesized that the combination of a HEHEHE-tag at the N terminus and a GGGC chelator at the C terminus of an affibody ...
Gentaur molecular products has all kinds of products like :search , Trevigen \ PAR Monoclonal Antibody Affinity Purified \ 4335-AMC-050 for more molecular products just contact us
Importance of herbal medicines have recently increased owing to rising interest in their health benefits. However, medicinal plant extracts are complex mixtures of phytochemicals that act synergistically or additively on specific and/or multiple molecular and cellular targets. Thus, it is difficult to examine the actual pharmacological roles of active compounds in plant extracts. This review describes a new strategy for isolating target compounds from plant extracts using immunoaffinity columns coupled with monoclonal antibodies (mAbs) against natural compounds. Through one-step purification using mAb-coupled immunoaffinity columns, we succeeded in preparing a knockout (KO) extract, which contains all components except the target compound. Furthermore, we investigated the pharmacological effects of the KO extract to reveal the actual effects of a bioactive compound in the crude extract. This approach may help determine the potential function of target compounds in herbal medicines.
Immunoaffinity Column Rack [CR1] - Intended use: R-Biopharm Rhône has designed a simple and easy to use racking system for performing immunoaffinity column analysis. Inserts are also available for the rack.General information: With its solid, chemical resistant structure, RBRs variable position immunaffinity column rack gives users a stable foundation to handle multiple analysis. Holding up
A polyhistidine tag, usually his6, attached to either the amino-terminus or carboxyl-terminus of an expressed recombinant protein enables affinity purification of that protein. Using the principles of immobilized metal affinity chromatography (IMAC), the his-tagged fusion protein binds to a metal chelate-coated substrate and can be eluted with imidazole. In addition to plant-plant proteomic applications, His-tagged proteins from bacterial or animal sources may be expressed in plants to avoid cross-contamination of the purified protein with other bacterial or animal proteins.1 Our HIS-Select nickel affinity system uses tetradentate chelated nickel for high selectivity bound to its support with a neutral spacer to minimize unwanted ionic interactions. HIS-select nickel affinity gel can bind about 15 mg protein/ml. The ligand has the same high capacity on highly crosslinked agarose for fast flow and high pressure chromatography, and in the EZview Red agarose format. HIS-select spin
Blurry bands ... - posted in SDS-PAGE and Western Blotting: Hello, Ive been running SDS Pages for a while ... lately I was doing some DNA affinity chromatography experiments and was running the elutions on a 16% gel. My particular interest is the identification of smaller proteins (around 10 - 20 kDa). I dont know why, but apparently the smaller proteins on my gel, appear as blurry bands (see attached file); thus giving really troubles for further MALDI analysis. Does anyone kno...
Purified proteins are often needed in the basic research laboratory and for diagnostic and therapeutic procedures. An effective technique for protein purification is affinity chromatography, which exploits a specific interaction between a protein and a complementary binding molecule. In this exercise, students isolate albumin from horse serum by affinity chromatography using a column matrix containing a reactive blue dye, which binds specifically to the albumin molecule. They then use electrophoresis to analyze the isolated protein in order to verify the effectiveness of the procedure.. Precast polyacrylamide gels can be ordered from Bio Rad, (800-424-6723) Their catalog number is 1611177.. ...
Affinity chromatography is an efficient method to isolate proteins by taking advantage of their affinities for specific molecules such as substrates, inhibitors, antigens, ligands, antibodies, and other interacting molecules including subunits. Nowadays, we take the effectiveness and excellence of this technology for granted. This essay will mainly cover the use of affinity chromatography based on my experience.
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(ID:10288) Development of an affinity chromatography purification of cell culture derived Vaccinia Virus VV after an initial host
Extensive research in the past two decades has led to the realization of Immunoglobulin-M (IgM) as a potential therapeutic and diagnostic agent. In order to fully exploit the potential of IgM, large quantities, in a highly pure and active form, must be available at low cost for performing clinical trials, characterization studies and quantitative-structure activity analyses. The complex physico-chemical properties, in particular its large size and labile nature renders downstream purification of IgM difficult. This review discusses the limitations and challenges associated with the current IgM purification strategies and proposes future directions for research. The uniqueness of affinity chromatography, specifically biomimetic affinity chromatography for protein purification is highlighted and its potential for IgM purification is discussed. © 2011 Elsevier Inc ...
A common structural feature of all cellular eukaryotic mRNAs (except for organelle mRNAs) analyzed to date is the presence of a 5′-terminal m7GpppN (where N is any nucleotide) structure, termed the cap (140). This structure is added early during RNA polymerase II-driven transcription and is required for several steps of mRNA biogenesis. Consistent with the diverse roles of the cap during gene expression, a number of cap binding proteins have been identified in the cytoplasm and nucleus (54, 140). The first-described, and best-characterized of these, is eukaryotic initiation factor-4E (eIF-4E), a 24-kDa polypeptide that has been cloned and characterized from a number of species. This protein shows strong binding specificity for methylated, cap structures of eukaryotic mRNAs (54).. We have developed an affinity chromatography procedure, called CAPture, that allows for the purification of mRNA via the cap structure (44). Previous to this, several laboratories had used antibodies directed against ...
Measure 1cm from bottom of strip and draw a line across Draw a cross in the centre of this line Attach a pin to the top of the chromatography paper, and p
The experiments in this thesis were performed to determine if novel uses of three fusion proteins could be established as a means of improving the protein purification process, and in particular, the elution step, thus resulting in the establishment of novel immunoaffinity purification methods. There are numerous fusion tags currently available for use as purification tools. Many of these current methods for protein purification require harsh elution steps, such as a low pH elution, which can be harmful to the protein. There are few purification methods that successfully purify protein, while maintaining a gentle pH environment for the protein. The goal was to employ a new in-house tag, and enhance two previously established tags, to optimize purification methods that would not require low pH elution steps. For each fusion tag in this work, there were three major aspects to the establishment of the purification method. The first aspect was the expression and purification by Nickel-IMAC of the ...
Amersham Biosciences Handbook. Affinity chromatography separates proteins on the basis of a reversible interaction between a protein (or group of proteins) and a specific ligand coupled to a chromatography matrix. The technique offers high selectivity, hence high resolution, and usually high capacity for the protein(s) of interest. Purification can be in the order of several thousand-fold and recoveries of active material are generally very high.. ...
affinity chromatography - posted in Protein and Proteomics: dear everyone who expert in protein purification I am confuse .... what is the differences between glutathione agarose and glutathione sepharose I am very grateful for somebody that can help me C U Erlia
Omnifit Chromatography Column Replacement Endpieces: Fixed Fixed endpiece; Bore size: 10mm; Operating pressure: 600 psi Omnifit Chromatography Column Replacement...
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This antibody was developed using β-estradiol-6-one-6-(O-carboxymethyloxime)-KLH conjugates as the immunogen and affinity purified with antigen-specific affinity chromatography. The antibody could be utilized for detection, quantization and affinity isolation and purification of estradiol in biofluid. ...
This simple guide will allow you to easily select the right basic buffer for the right protein purification strategy in order to facilitate your decision making.
보다 유용함이 밝혀졌다. 이 두가지 affinity media에 흡착된 효소를 분리 하는데 가장 적합한 elution 조건을 조사하였는데 KCI gradient( (0-1.OM)가 효소의 순도 및 수회율을 가장 높일 수 있는 적합한 방법이었다. 특히 Affi-gel Blue를 사용할 경우, KCI gradient로 효소를 용출시키기 전에 NAD-(15mM)로NAD+에 친화역을 갖는 효소들을 제거하는 것이 enzyme의 순도를 높이는데 매우 효과적이었다. 그 결과 glucose-6-phosphate dehydrogenase를 bakers yeast로 부터 기존의 간단한 정제 과정과 affinity chromatography를 병행한 방식을 샤용하여 분리 하였는데, affinity medium으로 Affi-gel Blue를 사용했을 때는 180배 정도, NADP+-agarose를 사용했을 때는 2,000배 정도로 정제 되었다. 대량으로 glucose-6-phosphate dehydrogenase를 정제하는 경우, Affi-gel Blue를 사용하던 효소의 순도는 NADP+-agarose보다 낮으나, 효소의 회수율은 ...
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Product Search Company Search Kit Search Gene Link specializes in complex modified and long oligos up to 250mer Advansta offers tools for protein purification analysis and characterization Free samples upon request Use these categories to find what you are looking for Custom Services Immunology related Services Antibody Affinity Isolation Antibody Characterization Testing Antibody Fab and F ab 2 Preparation Antibody Fragmentation Antibody Labeling Antibody Purification Antibody Quantitation Antigen Production ...
Recombinant human HBZ, fused to His-tag at C-terminus, was expressed in E.coli and purified by using conventional chromatography techniques.
The human and animal proteomes are area of large scientific interest for identifying desease biomarkers. Their proteomic approach is challenging because most of proteins of interest to be diagnosed are present at very low concentration in specialized cell types and biological fluids ...
Development of Novel Affinity Reagents. During the past fiscal year, the Office of Science and Technology Partnerships (OSTP) has continued to support an initia...
Recombinant human CDCA8 protein, fused to His-tag at N-terminus, was expressed in E.coli and purified by using conventional chromatography techniques (19-280aa).
Recombinant human RPE protein, fused to His-tag at N-terminus,was expressed in E. coli and purified byusing conventional chromatography techniques.
Protein purification, protein production - discover tips and basics for optimizing your rec. protein production and purification process.
a term sometimes used to denote methods of purification of substances by affinity chromatography in which a specific contaminant in a sample interacts with and is selectively retained by the adsorbent, especially as opposed to positive chromatography.. [...] ...
a vertical glass tube used in column chromatography; a mixture is poured in the top and washed through a stationary substance where components of the mixture are adsorbed selectively to form colored bands. ...
Experts share best practices for downstream purification, and how continuous purification methods can offer increased productivity and greater process reliability and reproducibility in the laboratory and when scaling protein purification processes for the production of commercial quantities ...
Identification, Production, and Use of Polyol-Responsive Monoclonal Antibodies for Immunoaffinity Chromatography / Nancy E. Thompson ; Katherine M. Foley ; Elizabeth S. Stalder ; And Richard R. ...
The ViraTrapTM AAV purification kit is designed for fast and efficient purification of all rAAV serotypres from AAV infected cell culture. Viral particles can be purified from cell culture of 5 to 6 T75 flasks per column. The viruses are first app
Our SeqPure PCR purification kit uses paramagnetic bead technology for rapid and efficient purification of PCR products. Click here to learn more!
A method for removing contaminant DNA in a sample containing a physiologically active protein, which comprises the following steps: 1) converting the sample containing a physiologically active protein into a neutral aqueous solution of low conductivity; and 2) removing the resulting particles.
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Most of MBLs PURIFICATION KITs accept 4℃ in elution step. However, in His tagged Protein PURIFICATION KIT, the purification efficiency is reduced to almost halve if you elute the epitope-tagged target protein at 4℃ . If you have to perform elution step at 4℃ , please incubate His-tag beads with Elution Peptide Solution at 4℃ overnight before elution ...
Dr. Srivatsan directs the neuroscience laboratory. She employs cellular and molecular techniques such as cell culture, immunocytochemistry, affinity chromatography, electrophoresis, ELISA, Western and Northern blots, spectrophotometry, light and fluorescence microscopy, real time imaging, calcium imaging, morphometry and microarray (gene chip) analysis in her research.. ...
Human prostatic acid phosphatase (PAP) isoenzymes, designated PAP-A and PAP-B, were isolated from human seminal plasma by sequential affinity chromatography on concanavalin A and L(+)-tartrate, a classic inhibitor of PAP. Both the major PAP-A and the minor PAP-B isoenzymes exhibited a similar molecular mass (100 and 105 kDa respectively), multiple pI values (5.05-5.35 and 5.05-5.12), and substrate and inhibitor specificity. Immunological characterization revealed that PAP-B possesses distinct antigenic determinants, in addition to the common sites shared with PAP-A. SDS/PAGE indicated that both isoenzymes are composed of two subunits of 50 kDa each. At high salt concentration, PAP-B dissociated completely into single subunits of 50 kDa, whereas PAP-A remained intact at 100 kDa. PAP-B was resolved by reverse-phase h.p.l.c. into three components, designated alpha, beta and gamma, each of 50 kDa, at a molar ratio of approx. 2:1:1. PAP-A contained a single component of molecular mass 50 kDa. The ...
During the mating reaction (fertilization) in the biflagellated alga, Chlamydomonas reinhardtii, mt+ and mt- gametes adhere to each other via their flagella and subsequently fuse to form quadriflagellated zygotes. In the studies reported here, we describe a monoclonal antibody directed against an mt+ flagellar surface molecule. The antibody blocks the adhesiveness of mt+ gametes, isolated mt+ flagella, and detergent extracts thereof. It has no effect on mt- gametes. Cyanogen bromide-activated Sepharose beads derivatized with the antibody bind only mt+ gametes; mt- gametes and mt+ and mt- vegetative cells are unreactive with the derivatized beads. The interaction of mt+ gametes with the beads is dynamic and cells continuously bind, detach, and rebind to the beads. Surprisingly, antibody-derivatized beads that have been incubated with mt+ gametes acquire the ability to bind mt- gametes. Moreover, extraction of the preincubated beads with detergents releases active mt+ adhesion molecules. The ...
This paper gives a capillary electrophoretic method for the separation of 15 urinary normal and modified nucleosides from cancer patients in less than 40 min. A 500 mmx50 mu m uncoated capillary column (437.5 mm to window) was used. The effects of the voltage and the sodium dodecyl sulfate (SDS) concentration in the buffer on the separation were studied. With reproducibilities of migration times better than 1.2% (R.S.D.) and determined concentrations better than 5-25%, depending on the concentrations of nucleosides in the urine, the analytical characteristics of the method were food. Using this developed method, the concentrations of 13 normal and modified nucleosides, extracted on a phenyl boronic acid affinity chromatography column, in 25 urines from patients of 14 kinds of cancer were determined. The levels (nmol/mol creatinine) of modified nucleosides in urines from cancer patients were increased as compared with those in normal urines. (C) 1998 Elsevier Science B.V ...
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Fragment-based drug discovery is an emerging process that has gained popularity in recent years. The process starts from small molecules called fragments. One major step in fragment-based drug discovery is fragment screening, which is a strategy to screen libraries of small molecules to find hits. The strategy in theory is more efficient than traditional high-throughput screening that works with larger molecules. As fragments intrinsically possess weak affinity to a target, detection techniques of high sensitivity to affinity are required for fragment screening. Furthermore, the use of different screening methods is necessary to improve the likelihood of success in finding suitable fragments. Since no single method can work for all types of screening, there is a demand for new techniques. The aim of this thesis is to introduce weak affinity chromatography (WAC) as a novel technique for fragment screening.. WAC is, as the name suggests, an affinity-based liquid chromatographic technique that ...
Thermostabilities of kanamycin nucleotidyltransferase and of its mutants that became thermostable, in the free state, because of single-amino-acid replacements were studied after immobilization of the enzymes on cyanogen bromide-activated Sephadex G-200 particles. Lys in place of Gln at position 102 decreased the thermostability of the immobilized enzyme, whereas replacement with other amino acids enhanced it. ...
This unit describes the purification of extracellular vitronectin from plasma or serum by using heparin‐affinity chromatography
Pseudobiospecific ligand l-histidine is an inexpensive, highly stable, non-toxic ligand explored successfully over the last twenty years for the purification of immunoglobulins in immobilised histidine ligand affinity chromatography. It is of great interest to know the molecular recognition sites of IgG to immobilized l-histidine. Here, we have used an in silico approach to explore the molecular recognition of l-histidine by IgG. We have assessed the feasible binding modes of histidine and its moieties at different sites of IgG and considered only those binding conformations which are exhibited via the imidazole ring NH group or any other OH donating group apart from the ones which are terminally conjugated with the support matrix. We categorised binding site into two categories; category I: inner binding groove and category II: surface binding groove and observed that the hinge region of IgG has most favourable binding pocket for l-histidine and histidyl moieties. Ser and Tyr residues on the hinge
Troponin-C Human produced from Human Cardiac Tissue has a mw of 18kDa and is purified using a combination of ion-exchange and affinity chromatography steps.
Periodical: Cuatrecasas, Pedro, M. Wilchek, and Christian B. Anfinsen. Selective Enzyme Purification by Affinity Chromatography. Proceedings of the National Academy of Sciences of the United States of America 61, 2 (October 1968): 636-643. Article. 8 Images ...
In this experiment Brooklyn and I prepared a 1/4 cup of water. Secondly we made a sample using filter paper. The third thing we did was we set up the filter paper in the cup of water making sure it was just touching the paper. Lastly we sketched a before and after sadly we forgot the before but this picture is after ...
This article is from Experimental & Molecular Medicine, volume 44.AbstractAptamers are synthetic, relatively short (e.g., 20-80 bases) RNA or ssDNA...
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PubMed comprises more than 30 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.
A method of purifying a recombinant protein from a solution, such as tissue culture fluid, containing gylcoproteins. The affinity of lectins for specific glycoproteins is assessed and used to select a
Since its founding in 1959-under the name Industrial Research-Ramp;D Magazine has served research scientists, engineers, and technical staff members at laboratories around the world. Ramp;D Magazine and www.rdmag.com provide timely, informative news and useful technical articles that broaden our readers knowledge of the research and development industry and improve the quality of their work. Our diverse content is delivered through Ramp;D Magazine,our print magazine published seven times per year, the Ramp;D Daily, our twice daily e-newsletter covering news of interest in all fields of Ramp;D and the annual Ramp;D 100 Awards, which celebrates technical advances in Ramp;D.
Recent technological advances in the way biologic therapeutics are purified may bring size-exclusion chromatography back into the modern purification process.
View Notes - Ch5AffinityChromatography from S 117 at Indiana. 5-1 Ch. 5 Affinity Chromatography for the Isolation of LDHObjectives Develop pipeting skills and practice dilution techniques Separate
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This application note demonstrates sample load optimization on a BioResolve SCX mAb Column, purification strategy using WFM-A, as well as a few examples of analyses that could be done on the charge-variant fractions in order to obtain useful information on the modification of the mAbs.
Fast protein liquid chromatography (FPLC) is a liquid chromatography technique for separation of protein molecules under pressure. It differs from high performance liquid chromatography in that the pressures are generally lower, around 435 to 580 psi, compared to 3000-to 5000 psi for an HPLC system.
DNA-protein interactions play an essential role in many regulatory mechanisms such as replication, transcription or translation and are responsible for the maintenance of the genome integrity. Isolation, identification and subsequent characterization of the biological function of DNA binding proteins propose insight into the pathological mechanisms that underlie various diseases. This review brings an overview of methods used for isolation and identification of DNA binding proteins (DNA-affinity chromatography coupled with quantitative proteomics and mass spectrometry) and subsequent methods for the characterization of their biological functions (fluorescence microscopy). Principles, advantages and disadvantages of individual methods are briefly discussed ...
Thank you for your interest in spreading the word about Biochemical Society Transactions.. NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.. ...
The tag strongly depends on your needs in your final experimental approach. Due to its small size and chemically inert nature, Strep-tag®II does generally not interfere with the folding or bioactivity of the recombinant protein.. The Twin-Strep-tag® enables the same mild and rapid purification as Strep-tag®II but, in addition, has an increased affinity for Strep-Tactin® and Strep-Tactin®XT which allows efficient purification (even in batch or directly from cell culture supernatants). With the Twin-Strep-tag® we introduce an avidity effect, which consequently reduces the off-rate of the total-tag and finally enhances the binding affinity to Strep-Tactin® as well as Strep-Tactin ®XT. After binding of the protein and changing to wash buffer the curve stays on a high level since a Twin-Strep-tag® fusion protein dissociates completely, only when both tags leave their binding pockets at the same time. If one tag dissociates it is kept in close proximity to its pocket by the other tag and is ...
Filtração em gel: separação por faixas de PM Figure 4.3. Gel Filtration Chromatography. A mixture of proteins in a small volume is applied to a column filled with porous beads. Because large proteins cannot enter the internal volume of the beads, they emerge sooner than do small ones Carga? (histonas = +) Cromatografia de troca iônica + - Figure 4.4. Ion-Exchange Chromatography. This technique separates proteins mainly according to their net charge Substrato? Cromatografia de afinidade Figure 4.5. Affinity Chromatography. Affinity chromatography of concanavalin A (shown in yellow) on a solid support containing covalently attached glucose residues (G). Colunas de material finamente dividido • • muito mais sítios de interação tempo "infinito" de purificação... Solução: HPLC High-Pressure Liquid Chromatography Figure 4.6. High-Pressure Liquid Chromatography (HPLC). Gel filtration by HPLC clearly defines the individual proteins because of its greater resolving power: (1) ...
For sale is are portions of an Amicon chromatography column. ID is between 20 and 22mm. Overall length just over 90Cm. Made of acryl
ProSep-vA chromatography media are designed to facilitate highly efficient, cost effective purification of monoclonal, polyclonal and engineered antibodies. Find MSDS or SDS, a COA, data sheets and more information.
Use Affi-Gel Blue Gel for rapid albumin removal, enzyme purification, and the separation and purification of plasma proteins inlcuding human serum complement.
Whatever the complexity or the scale of your purification process is, the BUCHI preparative chromatography systems are designed to fulfill your changing needs. Together with a broad range of high performance flash chromatography columns, we provide you the optimized solution suited to your purification workflow.
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.
dpG antibodies were fractionated on a cellulose-double-stranded DNA column. The flow-through fraction bound denatured DNA but not double-stranded DNA (dsDNA). The dpG-eluted fraction bound dsDNA preferentially. The results show that proteins can recognise dpG in DNA by different mechanisms, some involving DNA unwinding.. ...
In article ,199609231357.JAA11376 at mail.med.cornell.edu,, mf at aol.com (W) wrote: #hey netters: # Ive been working on a method to isolate proteins comprising a putative #complex using an antibody to a single known protein in the complex. Ive #prepared polyclonal antibodies to the target protein, then Ive affinity #purified these antibodies and coupled them to a protein A bed (using DMP #as a covalent coupling reagent). Then Ive taken extracts from appropriate #cells that we have previously shown express the target protein and passed #them across the bed to isolate the complex. At this point things have #gotten a little difficult. Eluates from the column have shown by western #that there is a very low yield of the target protein; moreover, when Ive #taken a small amount of the bed matrix and boiled in SDS-PAGE sample #buffer, I get the target protein in expected amounts. Taken together Ive #interpreted this data as an indication that the elution conditions Im #using (0.2N Acetic acid, ...
The core houses the inner frit, through which the eluent percolates and exits at the base of the column to a detector and hence to a fraction collector. The outer frit constitutes the column inlet, and consequently the sample has initially an extremely large area of stationary phase with which to interact. This renders the loading capacity of the radial flow column also very high. It is interesting to note, that as the solute progress radially through the stationary phase bed towards the center, the effective cross-sectional area of the column will become smaller. Consequently, the plate volume of the column will decrease (see Plate Theory and Extensions ) as the solute moves to the center which will result in the solute being concentrated. However, as the solute bands progressively decrease in concentration due to normal dispersion processes (see Dispersion in Chromatography Columns ), this counteracts the concentration effect from reduced bed cross-section and prevents the column packing from ...
58). Equation (58) shows that the minimum value of (H) is solely dependent on the column radius (r) and the thermodynamic properties of the solute/phase system. As opposed to the optimum velocity, the minimum value of (H) is not dependent on the solute diffusivity.. ...
Affinity purification coupled with mass spectrometry (AP-MS) is a widely used approach for the identification of protein-protein interactions. However, for any given protein of interest, determining which of the identified polypeptides represent bona fide interactors versus those that are background contaminants (for example, proteins that interact with the solid-phase support, affinity reagent or epitope tag) is a challenging task. The standard approach is to identify nonspecific interactions using one or more negative-control purifications, but many small-scale AP-MS studies do not capture a complete, accurate background protein set when available controls are limited. Fortunately, negative controls are largely bait independent. Hence, aggregating negative controls from multiple AP-MS studies can increase coverage and improve the characterization of background associated with a given experimental protocol. Here we present the contaminant repository for affinity purification (the CRAPome) and ...
A fluid purification system has cells whose purifying capability can be regenerated. Some of the cells are arranged in series to reach a high level of purification. An automatic valve network is controlled to cycle the cells in a way that levels the loads on each, thereby maximizing the service interval for replacing expired cells, enabling all of the cells to be replaced at the same time after having each contributing approximately equally to the purification load, and operated such that at any one time, at least one cell is regenerated so as to enable continuous up-time.
* PuraTreat R, the gas purification treatment for heavy-duty applications, , * High-efficiency purification process, , , , , , BASF now offers specialty amino acid salt formulations under the PuraTreat R...
A fusion tag, called DYKDDDDK and consisting of eight amino acids (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) including an enterokinase-cleavage site, was specifically designed for immunoaffinity chromatography. It allows elution under non-denaturing conditions. Several antibodies against this peptide ha...
Purify protease-free IgG from diverse biopharma sources (cells, sera, ascites) with minimal sample prep. Economical single-step albumin removal from serum.
Biomigas proprietary DNA binding systems that allow the high efficient reversible binding of DNA to the mini column while proteins and other impurities are removed by wash buffer. This kit is designed for fast and efficient purification of plasmid DNA fr
Escherichia coli IMP dehydrogenase (EC 1.2.1.14) was purified by affinity chromatography on immobilized nucleotides. The enzyme binds to agarose-bound 8-(6-aminohexyl)-AMP, N6-(6-aminohexyl)-AMP and 8-(8-amino-octyl)-IMP but not to immobilized NAD+ or Cibacron Blue F3G-A. AMP proved to be an effective eluent. A large-scale purification scheme in which 8-(6-aminohexyl)-AMP-agarose was used resulted in a homogeneous preparation of IMP dehydrogenase. The enzyme was also purified by immunoprecipitation with monospecific antisera. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, N-terminal amino acid analysis and tryptic finger-printing demonstrated that IMP dehydrogenase comprises identical subunits of mol.wt. 58000. Trypsin and Pronase cleave the 58000-mol.wt. subunit into peptides of mol.wts. 42000 and 14000, with a concomitant decrease in enzyme activity. These observations rationalize much of the contradictory data on the subunit composition of the enzyme found in the literature. ...
Research Corridor has published a new research study titled "Ion Exchange Chromatography Columns Market - Growth, Share, Opportunities, Competitive Analysis and Forecast, 2017 - 2025". The Ion Exchange Chromatography Columns market report studies current as well as future aspects of the Ion Exchange Chromatography Columns Market based upon factors such as market dynamics, key ongoing trends and segmentation analysis. Apart from the above elements, the Ion Exchange Chromatography Columns Market research report provides a 360-degree view of the Ion Exchange Chromatography Columns industry with geographic segmentation, statistical forecast and the competitive landscape.. Browse the complete report at http://www.researchcorridor.com/ion-exchange-chromatography-columns-market/. Geographically, the Ion Exchange Chromatography Columns Market report comprises dedicated sections centering on the regional market revenue and trends. The Ion Exchange Chromatography Columns market has been segmented on the ...
Rhizomucor miehei lipase (RML) is greatly hyperactivated (around 20- to 25-fold toward small substrates) in the presence of sucrose laurate. Hyperactivation appears to be an intramolecular process because it is very similar for soluble enzymes and covalently immobilized derivatives. The hyperactivated enzyme was immobilized (in the presence of sucrose laurate) on cyanogen bromide-activated Sepharose (very mild covalent immobilization through the amino terminal residue), on glyoxyl Sepharose (intense multipoint covalent immobilization through the region with the highest amount of Lys residues), and on different anion exchangers (by multipoint anionic exchange through the region with the highest density of negative charges). Covalent immobilization does not promote the fixation of the hyperactivated enzyme, but immobilization on Sepharose Q retains the hyperactivated enzyme even in the absence of a detergent. The hydrolysis of fish oils by these hyperactivated enzyme derivatives was sevenfold ...
A 195,000 mol wt Plasmodium falciparum protein and processing fragments derived from it have been purified by monoclonal antibody affinity chromatography. A polyvalent antiserum has been raised against the purified protein and used to identify the terminal processing products associated with the merozoite. Three unique fragments of 83,000, 42,000, and 19,000 mol wt are present and they represent the major surface antigens of P. falciparum merozoites. ...
The NF-kappaB/Rel transcription factors participate in the activation of immune system regulatory genes and viral early genes including the human immunodeficiency virus type 1 long terminal repeat. NF-kappaB/Rel proteins are coupled to inhibitory molecules, collectively termed IkappaB, which are responsible for cytoplasmic retention of NF-kappaB. Cell activation leads to the phosphorylation and degradation of IkappaBalpha, permitting NG-kappaB/Rel translocation to the nucleus and target gene activation. To further characterize the signaling events that contribute to IkappaBalpha phosphorylation, a kinase activity was isolated from Jurkat T cells that specifically interacted with IkappaBalpha in an affinity chromatography step and phosphorylated IkappaBalpha with high specificity in vitro. By using an in-gel kinase assay with recombinant IkappaBalpha as substrate, two forms of the kinase (43 and 38 kDa) were identified. Biochemical criteria and immunological cross-reactivity identified the kinase ...
The affinity of metal chelates for amino acids, such as histidine, is widely used in purifying proteins, most notably through six-histidine `tails. We have found that metal affinity interactions can also be applied to separation of single-stranded nucleic acids through interactions involving exposed purines. Here we describe a metal affinity precipitation method to resolve RNA from linear and plasmid DNA. A copper-charged copolymer of N-isopropyl acrylamide (NIPAM) and vinyl imidazole (VI) is used to purify plasmid from an alkaline lysate of E. coli. The NIPAM units confer reversible solubility on the copolymer while the imidazole chelates metal ions in a manner accessible to interaction with soluble ligands. RNA was separated from the plasmid by precipitation along with the polymer in the presence of 800 mM NaCl. Bound RNA could be recovered by elution with imidazole and separated from copolymer by a second precipitation step. RNA binding showed a strong dependence on temperature and on the ...
Fish oil made from menhaden (Brevoortia tyrannus) can be used as a dietary supplement for the presence of high levels of the long-chained omega-3 fatty acids, viz. epentaenoic and docosahexanoic. In this work, for the first time, two different multidimensional approaches were developed and compared, in terms of peak capacity, for triacylglycerol characterization. In particular, silver ion chromatography with a silver-ion column and non-aqueous reverse-phase liquid chromatography with a C18 column were tested in both comprehensive (stop-flow) and off-line modes. The use of mass spectra attained by atmospheric pressure chemical ionization for both LC approaches, and the fatty acids methyl esters profile of menhaden oil obtained by gas chromatography analysis, greatly supported the elucidation of the triacylglycerol content in menhaden oil. The off-line approach afforded a better separation and, thus, higher peak capacity to allow identifying and semiquantifying more than 250 triacylglycerols. Such a huge
Thank you for your interest in spreading the word about Biochemical Society Transactions.. NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.. ...

Affinity chromatography - WikipediaAffinity chromatography - Wikipedia

Weak affinity chromatography[edit]. Weak affinity chromatography[28] (WAC) is an affinity chromatography technique for affinity ... Immobilized metal ion affinity chromatography[edit]. Immobilized metal ion affinity chromatography (IMAC) is based on the ... Boronate affinity chromatography[edit]. Boronate affinity chromatography consists of using boronic acid or boronates to elute ... Lectin affinity chromatography is a form of affinity chromatography where lectins are used to separate components within the ...
more infohttps://en.wikipedia.org/wiki/Immunoaffinity_chromatography

Affinity chromatography | chemistry | BritannicaAffinity chromatography | chemistry | Britannica

... chromatography: Subsequent developments: A technique exhibiting great selectivity, affinity chromatography, was first described ... In chromatography: Retention mechanism. Affinity chromatography exploits this feature by binding a ligand with the desired ... In chromatography: Subsequent developments. A technique exhibiting great selectivity, affinity chromatography, was first ... interactive capability to a support such as a gel used in gel-filtration chromatography. The ligand retards a solute with the ...
more infohttps://www.britannica.com/science/affinity-chromatography

Affinity Chromatography and Biological Recognition - 1st EditionAffinity Chromatography and Biological Recognition - 1st Edition

Purchase Affinity Chromatography and Biological Recognition - 1st Edition. Print Book & E-Book. ISBN 9780121665807, ... Affinity Chromatography of Interferons-A Novel Application for Calmodulin-Sepharose. Dextran-Sepharose Affinity Chromatography ... Affinity Methods-Design and Development. IMA-Chromatography (Immobilized Ion Affinity Chromatography): Reflections of ... Recent Advances in High Performance Liquid Affinity Chromatography (HPLAC). High Performance Liquid Affinity Chromatography: ...
more infohttps://www.elsevier.com/books/affinity-chromatography-and-biological-recognition/chaiken/978-0-12-166580-7

Affinity Chromatography vs. Protein A/G PurificationAffinity Chromatography vs. Protein A/G Purification

Affinity chromatography and protein A/G purification are antibody purification methods to remove ligands from extracts. They ... Affinity chromatography. Affinity chromatography can be applied to a wide array of proteins and works by exploiting the binding ... Comparison of affinity chromatography and protein A/G purification. Both affinity chromatography and protein A/G purification ... Protein A/G affinity is a type of affinity chromatography that can be applied to antibodies. A and G proteins from ...
more infohttps://www.news-medical.net/life-sciences/Affinity-Chromatography-vs-Protein-AG-Purification.aspx

Immobilized gallium(III) affinity chromatography of phosphopeptides.  - PubMed - NCBIImmobilized gallium(III) affinity chromatography of phosphopeptides. - PubMed - NCBI

As a result of a systematic study, we propose the use of an immobilized metal affinity chromatography (IMAC) in a microtip ( ... Immobilized gallium(III) affinity chromatography of phosphopeptides.. Posewitz MC1, Tempst P. ...
more infohttps://www.ncbi.nlm.nih.gov/pubmed/10424175?dopt=Abstract

Affinity chromatography : methods and protocols (eBook, 2000) [WorldCat.org]Affinity chromatography : methods and protocols (eBook, 2000) [WorldCat.org]

Affinity chromatography : methods and protocols. [Pascal Bailon;] -- Annotation,p,Affinity chromatography is a powerful tool, ... This comprehensive collection of detailed affinity chromatography methods ... ... An overview of affinity chromatography --. Weak affinity chromatography --. Fluidized-bed recepter-affinity chromatography --. ... DNA affinity chromatography -- Affinity chromatography of pyrogens -- Western cross blot -- Affinity perfusion chromatography ...
more infohttp://www.worldcat.org/title/affinity-chromatography-methods-and-protocols/oclc/227334564

Lectin affinity chromatography of factor VIII - Miles Inc.Lectin affinity chromatography of factor VIII - Miles Inc.

The affinity of lectins for specific glycoproteins is assessed and used to select a ... Both groups showed that the secreted activity (further purified by affinity chromatography at Genentech) was able to correct ... Various other methods using affinity chromatography have been described for concentrating Factor VIII from plasma. Andersson, ... The affinity of lectins for specific glycoproteins is assessed and used to select a particular lectin specific for the ...
more infohttp://www.freepatentsonline.com/4981951.html

affinity chromatography: post #1affinity chromatography: post #1

affinity chromatography - posted in Protein and Proteomics: dear everyone who expert in protein purification I am confuse .... ...
more infohttp://www.protocol-online.org/forums/topic/4447-affinity-chromatography/

Affinity Chromatography Market - Analysis and Global Forecast to 2023 | MarketsandMarketsAffinity Chromatography Market - Analysis and Global Forecast to 2023 | MarketsandMarkets

The Affinity Chromatography is a type of liquid chromatography that uses a biologically related agent as a stationary phase to ... Biospecific ligand based consumables include protein A affinity, protein G affinity, protein L affinity, and lectin affinity; ... The Affinity Chromatography is a type of liquid chromatography that uses a biologically related agent as a stationary phase to ... and dye ligand-based affinity. High demand for protein A and immobilized metal affinity chromatography by pharmaceutical ...
more infohttps://www.marketsandmarkets.com/Market-Reports/affinity-chromatography-market-138402526.html

Gold Immunoprobes | Laboratory Affinity Chromatography Supplies | Spectrum ChemicalGold Immunoprobes | Laboratory Affinity Chromatography Supplies | Spectrum Chemical

SpectrumChemical.com carries a full line of Affinity Chromatography Sup Spectrum Chemical has a complete line of laboratory ... Chromatography Lab Supplies *Affinity Chromatography Columns and Accessories. *Gas Chromatography Columns and Accessories ...
more infohttps://www.spectrumchemical.com/OA_HTML/Supplies_Chromatography-Supplies_Affinity-Chromatography_Other.jsp?queryStr=J

Improved immobilized metal affinity chromatography for large-scale phosphoproteomics applications.  - PubMed - NCBIImproved immobilized metal affinity chromatography for large-scale phosphoproteomics applications. - PubMed - NCBI

Improved immobilized metal affinity chromatography for large-scale phosphoproteomics applications.. Ndassa YM1, Orsi C, Marto ... In principle, immobilized metal affinity chromatography (IMAC) represents an ideal enrichment method for phosphoproteomics. ...
more infohttps://www.ncbi.nlm.nih.gov/pubmed/17022650?dopt=Abstract

Affinity chromatographyAffinity chromatography

... is a method of separating biochemical mixtures and based on a highly specific ... Lectin affinity chromatography is a form of affinity chromatography where lectins are used to separate components within the ... affinity chromatography - Chromatography in which the immobile phase (bed material) has a specific biological affinity for the ... Immobilized metal ion affinity chromatography. Immobilized metal ion affinity chromatography (IMAC) is based on the specific ...
more infohttps://en.academic.ru/dic.nsf/enwiki/704970/49348

Cellufine ETclean Affinity Chromatography Media | amsbioCellufine ETclean Affinity Chromatography Media | amsbio

Cellufine ETclean Affinity Chromatography Media. Introduction The Cellufine ETclean is poly(ε-lysine) immobilized Cellufine ( ... The beads are a stable affinity beads that are resistant against the cleanup solutions, which include 0.2 M sodium hydroxide ... of the beads was estimated from calibration curves obtained by size exclusion chromatography. Pullulan and maltose were used ...
more infohttp://www.amsbio.com/cellufine-etclean-endotoxin-removal-media.aspx

Protein-protein interaction affinity chromatography of leukotriene C4 synthaseProtein-protein interaction affinity chromatography of leukotriene C4 synthase

A novel affinity chromatography purification for human leukotriene C4 synthase is described. It is based on a specific ... Protein-protein interaction affinity chromatography of leukotriene C4 synthase. Söderström, Mats Wallenberg Laboratory, ... Microsomal glutathione S-transferase was immobilized on NHS-activated Sepharose 4B and used as an affinity matrix. Microsomes ... Analyses of proteins specifically adsorbed to the affinity matrix revealed components with apparent molecular weights of 18, 37 ...
more infohttp://liu.diva-portal.org/smash/record.jsf?pid=diva2%3A658728

Protein Purification Using Affinity Chromatography | Thermo Fisher Scientific - ARProtein Purification Using Affinity Chromatography | Thermo Fisher Scientific - AR

Affinity Chromatography. Affinity chromatography is an effective technique for protein purification that often enables a single ... Affinity chromatography is a separation technique based on molecular conformation-molecules that "fit" one another bind ... Life Technologies offers a variety of products for affinity chromatography through its CaptureSelect® product portfolio. ... POROS® A and G affinity columns and CaptureSelect® affinity columns-operating instructions (PDF) ...
more infohttp://www.thermofisher.com/ar/en/home/references/protein-analysis-guide/affinity-chromatography.html

Protein Purification & Affinity Chromatography Resins & ColumnsProtein Purification & Affinity Chromatography Resins & Columns

... affinity chromatography resins for GST, 6X His and other common recombinant pr ... An extensive selection of affinity chromatography resins for protein purification featuring activated resins to generate your ... Purification & Chromatography * Affinity Purification Resins and Methods * Affinity Coupling * Avidin-Streptavidin Purification ... G-Biosciences Immobilized Monomeric Avidin Resin is designed for the simple affinity chromatography purifications of proteins, ...
more infohttps://www.gbiosciences.com/Protein-Research/Purification-Chromatography?page=8

Single-step purification of cyclotides using affinity chromatographySingle-step purification of cyclotides using affinity chromatography

affinity chromatography, cyclotides, cycloviolacin O2, immobilized IgG National Category Biochemistry and Molecular Biology ... Single-step purification of cyclotides using affinity chromatography. Uddin, Shaikh Jamal Uppsala University, Disciplinary ... Here, we describe the use of affinity chromatography for the purification of cyclotides using polyclonal IgG antibodies raised ... We demonstrate that single-step purification of cyclotides by affinity chromatography is possible but cross reactivity may ...
more infohttp://uu.diva-portal.org/smash/record.jsf?pid=diva2:1160014

Selective Enzyme Purification by Affinity Chromatography  (October 1968)Selective Enzyme Purification by Affinity Chromatography (October 1968)

Chromatography, Affinity Enzyme Inhibitors Exhibit Category: Molecular Engineering and Affinity Chromatography, 1959-1972 Box ... Selective Enzyme Purification by Affinity Chromatography Description: In this article, Cuatrecasas, Wilchek, and Anfinsen ... Selective Enzyme Purification by Affinity Chromatography. Proceedings of the National Academy of Sciences of the United States ... explain the general principles and potential applications of affinity chromatography, a protein purification technique that ...
more infohttps://profiles.nlm.nih.gov/ps/retrieve/ResourceMetadata/KKBBLF

Peptide Purification By Means of Metal Ion Affinity ChromatographyPeptide Purification By Means of Metal Ion Affinity Chromatography

... for purifying an objective protein and a polymer substrate for metal ion affinity chromatography. ; SOLUTION: The polymer ... Peptide Purification By Means of Metal Ion Affinity Chromatography. PROBLEM TO BE SOLVED: To provide an oligopeptide (tag) for ... purifying an objective protein and a polymer substrate for metal ion affinity chromatography. ; SOLUTION: The polymer substrate ...
more infohttps://www.ideaconnection.com/patents/10439-Peptide-Purification-By-Means-of-Metal-Ion-Affinity-C.html?c=6

Agarose bead size in affinity chromatography - Protein Expression and Purification - BioForumAgarose bead size in affinity chromatography - Protein Expression and Purification - BioForum

I have a quick question regarding agarose bead size in affinity chromatography, specifically nickel-coated beads for ... If I have a selection of the same column size with the same binding affinity, only differing in bead size (i.e. 34 µm or 165 µm ... Agarose bead size in affinity chromatography - posted in Protein Expression and Purification: Hello everybody, ... Chromatography machines? Started by wincel, 27 Jan 2016 chromatography, virus and 1 more... * 2 replies ...
more infohttp://www.protocol-online.org/forums/topic/30713-agarose-bead-size-in-affinity-chromatography/

Affinity Chromatography Archives - EY Laboratories, Inc.Affinity Chromatography Archives - EY Laboratories, Inc.

Copyright 2015 © EY Laboratories, Inc. All Rights Reserved. Reproduction of any materials from the site is strictly forbidden without permission.. ...
more infohttp://eylabs.com/labs/affinity-chromatography/

Affinity-chromatography Archives - EY Laboratories, Inc.Affinity-chromatography Archives - EY Laboratories, Inc.

Copyright 2015 © EY Laboratories, Inc. All Rights Reserved. Reproduction of any materials from the site is strictly forbidden without permission.. ...
more infohttp://eylabs.com/product-tag/affinity-chromatography-2/

Cellular membrane affinity chromatography (CMAC) in drug discovery from complex natural matricesCellular membrane affinity chromatography (CMAC) in drug discovery from complex natural matrices

artificial membrane stationary phase; bioassay; chromatography; frontal affinity chromatography; IAM.PC; immobilized protein; ... "Cellular membrane affinity chromatography (CMAC) in drug discovery from complex natural matrices." ADMET and DMPK, vol. 6, br. ... "Cellular membrane affinity chromatography (CMAC) in drug discovery from complex natural matrices." ADMET and DMPK 6, br. 3 ( ... Cellular membrane affinity chromatography (CMAC) allows the detection of secondary metabolites present in complex natural ...
more infohttps://hrcak.srce.hr/index.php?show=clanak&id_clanak_jezik=302774

Handbook of Affinity Chromatography, Second Edition 2nd edition | 9780824740573 | VitalSourceHandbook of Affinity Chromatography, Second Edition 2nd edition | 9780824740573 | VitalSource

Buy or Rent Handbook of Affinity Chromatography, Second Edition as an eTextbook and get instant access. With VitalSource, you ... Handbook of Affinity Chromatography, Second Edition 2nd Edition by David S. Hage and Publisher CRC Press. Save up to 80% by ...
more infohttps://www.vitalsource.com/products/handbook-of-affinity-chromatography-second-david-s-hage-v9780824751982
  • Affinity columns can be eluted by changing the ionic strength through a gradient. (academic.ru)
  • The following parameters can be determined using CMAC columns: binding affinity (Kd), association rate constant (kon), dissociation rate constant (koff) and the equilibrium constant for complex formation (K). This review summarizes the preparation steps and the use of CMAC columns in the drug discovery process of new potential drug leads present in complex natural matrices. (srce.hr)
  • In this report, the United States Affinity Chromatography Columns market is valued at USD XX million in 2016 and is expected to reach USD XX million by the end of 2022, growing at a CAGR of XX% between 2016 and 2022. (reportsnreports.com)
  • The Midwest with sales (volume), revenue (value), market share and growth rate of Affinity Chromatography Columns in these regions, from 2012 to 2022 (forecast). (reportsnreports.com)
  • Affinity chromatography can be used to purify and concentrate a substance from a mixture into a buffering solution, reduce the amount of unwanted substances in a mixture, identify the biological compounds binding to a particular substance, purify and concentrate an enzyme solution. (wikipedia.org)
  • The Affinity Chromatography is a type of liquid chromatography that uses a biologically related agent as a stationary phase to purify or analyze specific sample components. (marketsandmarkets.com)
  • Download this publication from Biotechnology Journal today to learn more about custom affinity chromatography development using camelid V H H antibody fragments, and their potential to purify highly pure and active biological therapeutics. (thermofisher.com)
  • Binding to the solid phase may be achieved by column chromatography whereby the solid medium is packed onto a column, the initial mixture run through the column to allow setting, a wash buffer run through the column and the elution buffer subsequently applied to the column and collected. (academic.ru)
  • As this isn't a "dumb" elution by time (as in size exclusion chromatography) but an elution with Imidazole in which I can determine the exact amount to add to specifically elute my protein of interest empirically, so the resolution should be the same no matter the bead size. (protocol-online.org)
  • Microsomes from 12-O-tetradecanoyl phorbol 13-acetate-treated human erythroleukemia cells were solubilized with taurocholic acid and applied on the affinity matrix at 0.1 M Mg2+ concentration. (diva-portal.org)