Molecular Weight: The sum of the weight of all the atoms in a molecule.Electrophoresis, Polyacrylamide Gel: Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.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 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.Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Gels: Colloids with a solid continuous phase and liquid as the dispersed phase; gels may be unstable when, due to temperature or other cause, the solid phase liquefies; the resulting colloid is called a sol.Chromatography, Gel: Chromatography on non-ionic gels without regard to the mechanism of solute discrimination.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.Electrophoresis, Gel, Two-Dimensional: Electrophoresis in which a second perpendicular electrophoretic transport is performed on the separate components resulting from the first electrophoresis. This technique is usually performed on polyacrylamide gels.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.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.Isoelectric Point: The pH in solutions of proteins and related compounds at which the dipolar ions are at a maximum.Kinetics: The rate dynamics in chemical or physical systems.Electrophoresis, Agar Gel: Electrophoresis in which agar or agarose gel is used as the diffusion medium.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)Electrophoresis, Capillary: A highly-sensitive (in the picomolar range, which is 10,000-fold more sensitive than conventional electrophoresis) and efficient technique that allows separation of PROTEINS; NUCLEIC ACIDS; and CARBOHYDRATES. (Segen, Dictionary of Modern Medicine, 1992)Body Weight: The mass or quantity of heaviness of an individual. It is expressed by units of pounds or kilograms.Electrophoresis, Disc: Electrophoresis in which discontinuities in both the voltage and pH gradients are introduced by using buffers of different composition and pH in the different parts of the gel column. The term 'disc' was originally used as an abbreviation for 'discontinuous' referring to the buffers employed, and does not have anything to do with the shape of the separated zones.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.Substrate Specificity: A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.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.Hydrogen-Ion Concentration: The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH = log 1/2[1/(H+)], where (H+) is the hydrogen ion concentration in gram equivalents per liter of solution. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)Chemistry: A basic science concerned with the composition, structure, and properties of matter; and the reactions that occur between substances and the associated energy exchange.Chemical Phenomena: The composition, conformation, and properties of atoms and molecules, and their reaction and interaction processes.Electrophoresis, Gel, Pulsed-Field: Gel electrophoresis in which the direction of the electric field is changed periodically. This technique is similar to other electrophoretic methods normally used to separate double-stranded DNA molecules ranging in size up to tens of thousands of base-pairs. However, by alternating the electric field direction one is able to separate DNA molecules up to several million base-pairs in length.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.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.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.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.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)Weight Loss: Decrease in existing BODY WEIGHT.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.Immunoelectrophoresis: A technique that combines protein electrophoresis and double immunodiffusion. In this procedure proteins are first separated by gel electrophoresis (usually agarose), then made visible by immunodiffusion of specific antibodies. A distinct elliptical precipitin arc results for each protein detectable by the antisera.Weight Gain: Increase in BODY WEIGHT over existing weight.Glycoproteins: Conjugated protein-carbohydrate compounds including mucins, mucoid, and amyloid glycoproteins.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.Sodium Dodecyl Sulfate: An anionic surfactant, usually a mixture of sodium alkyl sulfates, mainly the lauryl; lowers surface tension of aqueous solutions; used as fat emulsifier, wetting agent, detergent in cosmetics, pharmaceuticals and toothpastes; also as research tool in protein biochemistry.Bacterial Proteins: Proteins found in any species of bacterium.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.Peptide Fragments: Partial proteins formed by partial hydrolysis of complete proteins or generated through PROTEIN ENGINEERING techniques.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.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.Chromatography, High Pressure Liquid: Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.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)Liver: A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.Immunochemistry: Field of chemistry that pertains to immunological phenomena and the study of chemical reactions related to antigen stimulation of tissues. It includes physicochemical interactions between antigens and antibodies.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.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.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)Hot Temperature: Presence of warmth or heat or a temperature notably higher than an accustomed norm.Cell Line: Established cell cultures that have the potential to propagate indefinitely.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).DNA, Bacterial: Deoxyribonucleic acid that makes up the genetic material of bacteria.Microscopy, Electron: Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen.Isoenzymes: Structurally related forms of an enzyme. Each isoenzyme has the same mechanism and classification, but differs in its chemical, physical, or immunological characteristics.Antibodies, Monoclonal: Antibodies produced by a single clone of cells.Birth Weight: The mass or quantity of heaviness of an individual at BIRTH. It is expressed by units of pounds or kilograms.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.Protein Biosynthesis: The biosynthesis of PEPTIDES and PROTEINS on RIBOSOMES, directed by MESSENGER RNA, via TRANSFER RNA that is charged with standard proteinogenic AMINO ACIDS.Blood Protein Electrophoresis: Electrophoresis applied to BLOOD PROTEINS.Acrylic ResinsCross Reactions: Serological reactions in which an antiserum against one antigen reacts with a non-identical but closely related antigen.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.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 184.108.40.206.Electrophoresis, Starch Gel: Electrophoresis in which a starch gel (a mixture of amylose and amylopectin) is used as the diffusion medium.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)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.Electrophoresis, Microchip: A highly miniaturized version of ELECTROPHORESIS performed in a microfluidic device.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.Affinity Labels: Analogs of those substrates or compounds which bind naturally at the active sites of proteins, enzymes, antibodies, steroids, or physiological receptors. These analogs form a stable covalent bond at the binding site, thereby acting as inhibitors of the proteins or steroids.Cell Membrane: The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.Spectrophotometry: The art or process of comparing photometrically the relative intensities of the light in different parts of the spectrum.Carrier Proteins: Transport proteins that carry specific substances in the blood or across cell membranes.Swine: Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA).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.Plants: Multicellular, eukaryotic life forms of kingdom Plantae (sensu lato), comprising the VIRIDIPLANTAE; RHODOPHYTA; and GLAUCOPHYTA; all of which acquired chloroplasts by direct endosymbiosis of CYANOBACTERIA. They are characterized by a mainly photosynthetic mode of nutrition; essentially unlimited growth at localized regions of cell divisions (MERISTEMS); cellulose within cells providing rigidity; the absence of organs of locomotion; absence of nervous and sensory systems; and an alternation of haploid and diploid generations.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.Hydrolysis: The process of cleaving a chemical compound by the addition of a molecule of water.Genes, Bacterial: The functional hereditary units of BACTERIA.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.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.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.Protease Inhibitors: Compounds which inhibit or antagonize biosynthesis or actions of proteases (ENDOPEPTIDASES).Genes: A category of nucleic acid sequences that function as units of heredity and which code for the basic instructions for the development, reproduction, and maintenance of organisms.Electrophoresis, Cellulose Acetate: Electrophoresis in which cellulose acetate is the diffusion medium.Binding Sites: The parts of a macromolecule that directly participate in its specific combination with another molecule.Recombinant Proteins: Proteins prepared by recombinant DNA technology.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).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.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.Viral Proteins: Proteins found in any species of virus.AcrylatesDensitometry: The measurement of the density of a material by measuring the amount of light or radiation passing through (or absorbed by) the material.Epitopes: Sites on an antigen that interact with specific antibodies.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.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.Cell-Free System: A fractionated cell extract that maintains a biological function. A subcellular fraction isolated by ultracentrifugation or other separation techniques must first be isolated so that a process can be studied free from all of the complex side reactions that occur in a cell. The cell-free system is therefore widely used in cell biology. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p166)Acrylamides: Colorless, odorless crystals that are used extensively in research laboratories for the preparation of polyacrylamide gels for electrophoresis and in organic synthesis, and polymerization. Some of its polymers are used in sewage and wastewater treatment, permanent press fabrics, and as soil conditioning agents.Immunoelectrophoresis, Two-Dimensional: Immunoelectrophoresis in which a second electrophoretic transport is performed on the initially separated antigen fragments into an antibody-containing medium in a direction perpendicular to the first electrophoresis.Peptide Hydrolases: Hydrolases that specifically cleave the peptide bonds found in PROTEINS and PEPTIDES. Examples of sub-subclasses for this group include EXOPEPTIDASES and ENDOPEPTIDASES.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.Cell Fractionation: Techniques to partition various components of the cell into SUBCELLULAR FRACTIONS.TritiumAlcohol Oxidoreductases: A subclass of enzymes which includes all dehydrogenases acting on primary and secondary alcohols as well as hemiacetals. They are further classified according to the acceptor which can be NAD+ or NADP+ (subclass 1.1.1), cytochrome (1.1.2), oxygen (1.1.3), quinone (1.1.5), or another acceptor (1.1.99).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)Immunosorbent Techniques: Techniques for removal by adsorption and subsequent elution of a specific antibody or antigen using an immunosorbent containing the homologous antigen or antibody.Flavobacterium: A genus of gram-negative, aerobic, rod-shaped bacteria widely distributed in SOIL and WATER. Its organisms are also found in raw meats, MILK and other FOOD, hospital environments, and human clinical specimens. Some species are pathogenic in humans.Ammonium Sulfate: Sulfuric acid diammonium salt. It is used in CHEMICAL FRACTIONATION of proteins.Peptide Biosynthesis: The production of PEPTIDES or PROTEINS by the constituents of a living organism. The biosynthesis of proteins on RIBOSOMES following an RNA template is termed translation (TRANSLATION, GENETIC). There are other, non-ribosomal peptide biosynthesis (PEPTIDE BIOSYNTHESIS, NUCLEIC ACID-INDEPENDENT) mechanisms carried out by PEPTIDE SYNTHASES and PEPTIDYLTRANSFERASES. Further modifications of peptide chains yield functional peptide and protein molecules.Methods: A series of steps taken in order to conduct research.Sequence Homology, Nucleic Acid: The sequential correspondence of nucleotides in one nucleic acid molecule with those of another nucleic acid molecule. Sequence homology is an indication of the genetic relatedness of different organisms and gene function.Blood Proteins: Proteins that are present in blood serum, including SERUM ALBUMIN; BLOOD COAGULATION FACTORS; and many other types of proteins.Muscles: Contractile tissue that produces movement in animals.Drug Stability: The chemical and physical integrity of a pharmaceutical product.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.Enzyme Precursors: Physiologically inactive substances that can be converted to active enzymes.Plant Proteins: Proteins found in plants (flowers, herbs, shrubs, trees, etc.). The concept does not include proteins found in vegetables for which VEGETABLE PROTEINS is available.Polymerase Chain Reaction: In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.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.Fabaceae: The large family of plants characterized by pods. Some are edible and some cause LATHYRISM or FAVISM and other forms of poisoning. Other species yield useful materials like gums from ACACIA and various LECTINS like PHYTOHEMAGGLUTININS from PHASEOLUS. Many of them harbor NITROGEN FIXATION bacteria on their roots. Many but not all species of "beans" belong to this family.Cytosol: Intracellular fluid from the cytoplasm after removal of ORGANELLES and other insoluble cytoplasmic components.MercaptoethanolGlycosylation: 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.Sequence Analysis, DNA: A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.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.Autoradiography: The making of a radiograph of an object or tissue by recording on a photographic plate the radiation emitted by radioactive material within the object. (Dorland, 27th ed)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.Plants, Medicinal: Plants whose roots, leaves, seeds, bark, or other constituent parts possess therapeutic, tonic, purgative, curative or other pharmacologic attributes, when administered to man or animals.Nucleic Acid Denaturation: Disruption of the secondary structure of nucleic acids by heat, extreme pH or chemical treatment. Double strand DNA is "melted" by dissociation of the non-covalent hydrogen bonds and hydrophobic interactions. Denatured DNA appears to be a single-stranded flexible structure. The effects of denaturation on RNA are similar though less pronounced and largely reversible.Denaturing Gradient Gel Electrophoresis: Electrophoresis in which various denaturant gradients are used to induce nucleic acids to melt at various stages resulting in separation of molecules based on small sequence differences including SNPs. The denaturants used include heat, formamide, and urea.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.Immunologic Techniques: Techniques used to demonstrate or measure an immune response, and to identify or measure antigens using antibodies.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.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.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.Nucleic Acid Hybridization: Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503)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.Serine Endopeptidases: Any member of the group of ENDOPEPTIDASES containing at the active site a serine residue involved in catalysis.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.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.Ribonucleases: Enzymes that catalyze the hydrolysis of ester bonds within RNA. EC 3.1.-.Pronase: A proteolytic enzyme obtained from Streptomyces griseus.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.Endopeptidases: A subclass of PEPTIDE HYDROLASES that catalyze the internal cleavage of PEPTIDES or PROTEINS.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.Nucleoproteins: Proteins conjugated with nucleic acids.Proteomics: The systematic study of the complete complement of proteins (PROTEOME) of organisms.Phosphorus Radioisotopes: Unstable isotopes of phosphorus that decay or disintegrate emitting radiation. P atoms with atomic weights 28-34 except 31 are radioactive phosphorus isotopes.RNA, Viral: Ribonucleic acid that makes up the genetic material of viruses.Mass Spectrometry: An analytical method used in determining the identity of a chemical based on its mass using mass analyzers/mass spectrometers.Time Factors: Elements of limited time intervals, contributing to particular results or situations.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.SepharoseEdetic 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.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)Sulfur Radioisotopes: Unstable isotopes of sulfur that decay or disintegrate spontaneously emitting radiation. S 29-31, 35, 37, and 38 are radioactive sulfur isotopes.Rosaniline Dyes: Compounds that contain the triphenylmethane aniline structure found in rosaniline. Many of them have a characteristic magenta color and are used as COLORING AGENTS.GlucosamineKininogens: Endogenous peptides present in most body fluids. Certain enzymes convert them to active KININS which are involved in inflammation, blood clotting, complement reactions, etc. Kininogens belong to the cystatin superfamily. They are cysteine proteinase inhibitors. HIGH-MOLECULAR-WEIGHT KININOGEN; (HMWK); is split by plasma kallikrein to produce BRADYKININ. LOW-MOLECULAR-WEIGHT KININOGEN; (LMWK); is split by tissue kallikrein to produce KALLIDIN.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.Polyethylene Glycols: Polymers of ETHYLENE OXIDE and water, and their ethers. They vary in consistency from liquid to solid depending on the molecular weight indicated by a number following the name. They are used as SURFACTANTS, dispersing agents, solvents, ointment and suppository bases, vehicles, and tablet excipients. Some specific groups are NONOXYNOLS, OCTOXYNOLS, and POLOXAMERS.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.Cations, Divalent: Positively charged atoms, radicals or groups of atoms with a valence of plus 2, which travel to the cathode or negative pole during electrolysis.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)Erythrocytes: Red blood cells. Mature erythrocytes are non-nucleated, biconcave disks containing HEMOGLOBIN whose function is to transport OXYGEN.Reticulocytes: Immature ERYTHROCYTES. In humans, these are ERYTHROID CELLS that have just undergone extrusion of their CELL NUCLEUS. They still contain some organelles that gradually decrease in number as the cells mature. RIBOSOMES are last to disappear. Certain staining techniques cause components of the ribosomes to precipitate into characteristic "reticulum" (not the same as the ENDOPLASMIC RETICULUM), hence the name reticulocytes.Electrophoresis, Paper: Electrophoresis in which paper is used as the diffusion medium. This technique is confined almost entirely to separations of small molecules such as amino acids, peptides, and nucleotides, and relatively high voltages are nearly always used.Methionine: A sulfur-containing essential L-amino acid that is important in many body functions.Pseudomonas: A genus of gram-negative, aerobic, rod-shaped bacteria widely distributed in nature. Some species are pathogenic for humans, animals, and plants.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.Catalysis: The facilitation of a chemical reaction by material (catalyst) that is not consumed by the reaction.Proteome: The protein complement of an organism coded for by its genome.Glycoside HydrolasesPregnancy: The status during which female mammals carry their developing young (EMBRYOS or FETUSES) in utero before birth, beginning from FERTILIZATION to BIRTH.Native Polyacrylamide Gel Electrophoresis: Polyacrylamide gel electrophoresis under conditions in which the components, such as PROTEINS, being separated can remain in their naturally folded state.Metalloendopeptidases: ENDOPEPTIDASES which use a metal such as ZINC in the catalytic mechanism.DNA Restriction Enzymes: Enzymes that are part of the restriction-modification systems. They catalyze the endonucleolytic cleavage of DNA sequences which lack the species-specific methylation pattern in the host cell's DNA. Cleavage yields random or specific double-stranded fragments with terminal 5'-phosphates. The function of restriction enzymes is to destroy any foreign DNA that invades the host cell. Most have been studied in bacterial systems, but a few have been found in eukaryotic organisms. They are also used as tools for the systematic dissection and mapping of chromosomes, in the determination of base sequences of DNAs, and have made it possible to splice and recombine genes from one organism into the genome of another. EC 3.21.1.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.Antigen-Antibody Complex: The complex formed by the binding of antigen and antibody molecules. The deposition of large antigen-antibody complexes leading to tissue damage causes IMMUNE COMPLEX DISEASES.Collodion: A nitrocellulose solution in ether and alcohol. Collodion has a wide range of uses in industry including applications in the manufacture of photographic film, in fibers, in lacquers, and in engraving and lithography. In medicine it is used as a drug solvent and a wound sealant.RNA: A polynucleotide consisting essentially of chains with a repeating backbone of phosphate and ribose units to which nitrogenous bases are attached. RNA is unique among biological macromolecules in that it can encode genetic information, serve as an abundant structural component of cells, and also possesses catalytic activity. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)Culture Media: Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN.Nucleic Acid Conformation: The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape.Rotavirus: A genus of REOVIRIDAE, causing acute gastroenteritis in BIRDS and MAMMALS, including humans. Transmission is horizontal and by environmental contamination. Seven species (Rotaviruses A thru G) are recognized.Kidney: Body organ that filters blood for the secretion of URINE and that regulates ion concentrations.Gene Expression: The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.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.Restriction Mapping: Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.Poly A: A group of adenine ribonucleotides in which the phosphate residues of each adenine ribonucleotide act as bridges in forming diester linkages between the ribose moieties.Deoxyribonucleases: Enzymes which catalyze the hydrolases of ester bonds within DNA. EC 3.1.-.Antigens, Bacterial: Substances elaborated by bacteria that have antigenic activity.Dimethyl Suberimidate: The methyl imidoester of suberic acid used to produce cross links in proteins. Each end of the imidoester will react with an amino group in the protein molecule to form an amidine.Fluorescent Antibody Technique: Test for tissue antigen using either a direct method, by conjugation of antibody with fluorescent dye (FLUORESCENT ANTIBODY TECHNIQUE, DIRECT) or an indirect method, by formation of antigen-antibody complex which is then labeled with fluorescein-conjugated anti-immunoglobulin antibody (FLUORESCENT ANTIBODY TECHNIQUE, INDIRECT). The tissue is then examined by fluorescence microscopy.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).Heparin, Low-Molecular-Weight: Heparin fractions with a molecular weight usually between 4000 and 6000 kD. These low-molecular-weight fractions are effective antithrombotic agents. Their administration reduces the risk of hemorrhage, they have a longer half-life, and their platelet interactions are reduced in comparison to unfractionated heparin. They also provide an effective prophylaxis against postoperative major pulmonary embolism.Sulfhydryl Compounds: Compounds containing the -SH radical.Sulfhydryl Reagents: Chemical agents that react with SH groups. This is a chemically diverse group that is used for a variety of purposes. Among these are enzyme inhibition, enzyme reactivation or protection, and labelling.Protein PrecursorsUrea: A compound formed in the liver from ammonia produced by the deamination of amino acids. It is the principal end product of protein catabolism and constitutes about one half of the total urinary solids.Oxidoreductases: The class of all enzymes catalyzing oxidoreduction reactions. The substrate that is oxidized is regarded as a hydrogen donor. The systematic name is based on donor:acceptor oxidoreductase. The recommended name will be dehydrogenase, wherever this is possible; as an alternative, reductase can be used. Oxidase is only used in cases where O2 is the acceptor. (Enzyme Nomenclature, 1992, p9)
Proteins are introduced into an Immobilized pH gradient gel composed of polyacrylamide, starch, or agarose where a pH gradient ... in which proteins are first separated by their pI and then further separated by molecular weight through SDS-PAGE. According to ... Microchip based electrophoresis is a promising alternative to capillary electrophoresis since it has the potential to provide ... The method is applied particularly often in the study of proteins, which separate based on their relative content of acidic and ...
"Electrophoresis A Guide to Polyacrylamide Gel Electrophoresis and Detection" (PDF). Bio-Rad. "Protein Molecular Weight Markers ... Examples of these characteristics include affinity tags and molecular weights which are uniformly positioned relative to each ... One of the most common uses for molecular-weight size markers is in gel electrophoresis. The purpose of gel electrophoresis is ... Molecular-weight size markers are most commonly used in SDS-polyacrylamide gel electrophoresis and western blotting. With all ...
Another method that is common is polyacrylamide gel electrophoresis (PAGE). Both methods require amplification of all loci of ... CE is advantageous over PAGE in that molecular weight measurements like mass spectrometry can be used with analytes, whereas ... Examination of relatives of individuals thought to have sporadic ataxia can often reveal enough family history to identify a ... Molecular genetic testing of SCAs must be able to differentiate samples with the pathogenic allele from those without and be ...
"The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis". The Journal of ... Double-stranded DNA fragments naturally behave as long rods, so their migration through the gel is relative to their size or, ... Gel electrophoresis can also be used for separation of nanoparticles. Gel electrophoresis uses a gel as an anticonvective ... by native gel electrophoresis, by preparative gel electrophoresis (QPNC-PAGE), or by 2-D electrophoresis. Characterization ...
The acronym expands to "sodium dodecyl sulfate-polyacrylamide gel electrophoresis." Ninfa, Alexander; Ballou, David; Benore, ... The association of SDS molecules with protein molecules imparts an associated negative charge to the molecular aggregate formed ... CS1 maint: Uses authors parameter (link) [non-primary source needed] Islam, M. F. (2003). "High Weight Fraction Surfactant ... Bales, Barney L.; Messina, Luis; Vidal, Arwen; Peric, Miroslav & Nascimento, Otaciro Rangel (1998). "Precision Relative ...
Issaq, J. Haleem & T. D. Veenstra (2008). "Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE): advances and ... This dimension separates the protein according to its molecular weight. Once this step is completed in-gel digestion occurs. In ... The ratio of their peak intensities corresponds to the relative abundance ratio of the peptides (and proteins). The most ... The first method fractionates whole proteins via two-dimensional gel electrophoresis. The first-dimension of 2D gel is ...
... is independent of molecular weight during electrophoresis. The gel matrix is therefore responsible for the separation of DNA by ... Gel electrophoresis Immunodiffusion, Immunoelectrophoresis SDD-AGE Northern blot SDS-polyacrylamide gel electrophoresis ... and the DNA fragments can be extracted from the gel with relative ease. Most agarose gels used are between 0.7 - 2% dissolved ... Agarose gel electrophoresis is a method of gel electrophoresis used in biochemistry, molecular biology, genetics, and clinical ...
Hjertén S (1963). ""Molecular-sieve" electrophoresis in cross-linked polyacrylamide gels". Journal of Chromatography A. 11: 66- ... As both low and high molecular weight (metal) proteins and their respective structures can be analysed in a single run, a ... The relative biochemical impact of an applied metal-containing compound (metal-based drug) is depending on its dose, ... "Gel composition in gels for submurged gel electrophoresis". Patent US5458760 (A). Westermeier R (2016). Electrophoresis in ...
... one can make conclusions about the relative molecular weight of the proteins, where the length of the gel is determined by the ... It is common to run molecular weight size markers of known molecular weight in a separate lane in the gel to calibrate the gel ... Polyacrylamide gels are composed of a stacking gel and separating gel. Stacking gels have a higher porosity relative to the ... September 1967). "Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels". Biochem ...
... the ideal fragment lengths of the DNA for sequencing the fragments are size separated by polyacrylamide gel electrophoresis ( ... The high-molecular-weight DNA, often several megabase pairs long, is sonicated to break the DNA double-strands at random ... used DNA nanoball sequencing to sequence the genomes of a family of four relatives and were able to identify SNPs that may be ... PAGE). The DNA of the suitable size range is purified by gel extraction, resulting in DNA with lengths within a narrow range ( ...
Nature Methods 3(5): 410 article Moore, J. C. (1964) Gel Permeation Chromatography. 1. A New Method for Molecular Weight ... Lathe, GH and Ruthven, CR (1956) The separation of substances and estimation of their relative molecular sizes by the use of ... Either technique should not be confused with gel electrophoresis, where an electric field is used to "pull" or "push" molecules ... other gels with size fractionation properties include agarose and polyacrylamide. A short review of these developments has ...
DNA by restriction enzymes yields specific fragments that can be separated using polyacrylamide gel electrophoresis, thus ... List of homing endonuclease cutting sites List of restriction enzyme cutting sites Molecular weight size marker Restriction map ... A DNA map by restriction digest can also be generated that can give the relative positions of the genes. The different lengths ... Clark DP (2005). Molecular biology. Amsterdam: Elsevier Academic Press. ISBN 0-12-175551-7. Goodsell DS (2002). "The molecular ...
... which is used for separation of proteins by 2-D gel polyacrylamide gel electrophoresis. Buffer solutions also play a key role ... Relative stabilization of methylammonium ions thus decreases with the number of methyl groups explaining the order of water ... The value of pKa also depends on molecular structure of the acid in many ways. For example, Pauling proposed two rules: one for ... For substances in solution the isoelectric point (pI) is defined as the pH at which the sum, weighted by charge value, of ...
... which is used for separation of proteins by 2-D gel polyacrylamide gel electrophoresis. ... The value of pKa also depends on molecular structure of the acid in many ways. For example, Pauling proposed two rules: one for ... For substances in solution the isoelectric point (pI) is defined as the pH at which the sum, weighted by charge value, of ... Relative stabilization of methylammonium ions thus decreases with the number of methyl groups explaining the order of water ...
DNA by restriction enzymes yields specific fragments that can be separated using polyacrylamide gel electrophoresis, thus ... "European Molecular Biology Laboratory - Hamburg. Retrieved 2008-06-07.. *^ Russell DW, Sambrook J (2001). Molecular cloning: a ... A DNA map by restriction digest can also be generated that can give the relative positions of the genes. The different ... Molecular weight size marker. *Restriction map. *Star activity. *CRISPR. *CAS9. References. *^ Roberts RJ (Nov 1976). " ...
For example, before the advent of DNA gel electrophoresis (agarose or polyacrylamide), the size of DNA molecules was typically ... The northern blot is used to study the expression patterns of a specific type of RNA molecule as relative comparison among a ... Gel electrophoresis is one of the principal tools of molecular biology. The basic principle is that DNA, RNA, and proteins can ... Molecular biology /məˈlɛkjʊlər/ is the branch of biology that concerns the molecular basis of biological activity in and ...
... as indicated by the shifts in molecular weights of the glycoconjugates relative to the molecular weight of the unconjugated ... 12). Following conjugation, the samples were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). ... In order to dissect, at a molecular level, the binding of mAb 4C7 to Bp and Bm OAg, we employed ad hoc NMR techniques aimed to ... a molecular view achievable by using NMR spectroscopy and molecular modelling. Chem. Open 5, 274-296 (2016). ...
On SDS-polyacrylamide gel electrophoresis both PGs behave like proteins of the relative molecular weight of approximately ... We then used 14-3-3-stimulated ExoS modification of vH-Ras to screen a collection of low-molecular-weight compounds selected to ... Molecular modelling was undertaken using both docking and molecular dynamics. The (R)-and (S)-enantiomers of Flu-AM1 inhibited ... in the field of molecular modelling. More specifically, molecular docking was investigated as a tool for reproduction of ligand ...
TBP 2 binds transferrin after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroblotting, but TBP 1 does not ... Analogous higher- and lower-molecular-weight proteins associated with transferrin binding have been found in N. gonorrhoeae and ... The relative contributions of these two proteins to the binding reaction observed with intact cells and to iron uptake are ...
SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting were used to perform an analysis of the molecular weights ... The relative abundance of NHE-1 protein was not significantly different between thymic lymphocytes from nine SHR and nine WKY ... Both sets of antibodies recognized a protein band at about 110 kD (Fig 1⇓), the reported molecular weight of NHE-1.10 ... Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: ...
Analytical sodium dodecyl sulfate-polyacrylamide gel electrophoresis of human spinal cord components followed by immunoblots ... actually synthesize a surface antigen precipitated by 10E5 antibody as a major band with 92,000 relative molecular weight. Our ... Both alpha-(alpha-ETF, 32,000 molecular weight [mol wt]) and beta-subunits (beta-ETF, 27,000 mol wt) were nuclear-coded, and ... By immunoblotting, GPS reacted primarily with the 28,000 molecular weight (mol wt) monomer but also the 24,000 mol wt and ...
... and dextran using different molecular weights (40, 70, and 100 kDa) and mixing ratios was studied. This study includes the ... The covalent attachment of dextran to WPI was confirmed using sodium-dodecyl-sulfate-polyacrylamide gel-electrophoresis with ... relative humidity, 60 °C, 48 h). ... The impact of the molecular weight of dextran on formation of ... The impact of the molecular weight of dextran on formation of whey protein isolate (WPI)-dextran conjugates in fibers produced ...
Electrophoresis was performed at 4°C on polyacrylamide gel in PFO containing running buffer [25 mm Tris, 192 mm glycine and 0.5 ... Because the molecular weights of P2X2 and P2X5 subunits are similar, we used P2X subunits fused to the YC3.1 cameleon protein ( ... loading buffer containing NaPFO and separated by electrophoresis as described above. Relative migration of protein standards ... In both conditions (P2X5-Myc:P2X2-Cam or P2X2-Myc:P2X5-Cam), a single high molecular weight form, larger than the homotrimeric ...
The 32P-labeled proteins were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the 32P content was ... When the reaction mixture contained EGTA and no added Ca2+, 32P was incorporated into two proteins with molecular weights of ... visualized by autoradiography, and the relative intensities of specific bands were quantitated. ... Mr proteins and resulted in the phosphorylation of a major protein whose molecular weight (64,000 Mr) is identical to that of ...
Viral preparations were subjected to polyacrylamide gel electrophoresis (PAGE) followed by silver staining and the molecular ... followed by silver staining and the molecular weight and relative intensity of the viral capsid proteins was analyzed. The ... Purity of viral preparation: Viral preparations were subjected to polyacrylamide gel electrophoresis (PAGE) ... weight and relative intensity of the viral capsid proteins was analyzed. The abundance of viral capsid proteins as a fraction ...
... pooled on the basis of the relative molecular weight of the products as determined by SDS-polyacrylamide gel electrophoresis ( ... preparative SDS-polyacrylamide gel (16 cm by 20 cm by 1.5 mm) and electrophoresed at 20 mA for 1.5 hours. The resolved proteins ... 2 Molecular Biology Institute, Divisions of 5 Howard Hughes Medical Institute, University of California Los Angeles School of ... B) SDS-PAGE ofM. tuberculosis proteins from isoelectric focusing fraction 4 (10). Molecular size markers are in the left lane, ...
... by polyacrylamide gel electrophoresis and use of isoelectrofocusing gels in affected subjects revealed normal molecular weight ... The relative increase in proapolipoprotein AI was similar to that observed in patients with increased catabolism of HDL ... Molecular weight standards were applied on each gel to determine the size of the hybridized fragments. The gels were exposed on ... and standard molecular weight markers (low-molecular weight standards, Bio-Rad) were loaded in a separate well. Each probands ...
During electrophoresis in SDS-polyacrylamide gels there is a linear relationship between the logarithm of the molecular weight ... and the relative distance of the migration of the SDS-polypeptide micelle.. To calculate the molecular weight of an unknown ...
... made in HEp-2 cells infected with herpes simplex virus type 1 by high-resolution polyacrylamide gel electrophoresis revealed ... These 47 polypeptides can account for 75% of the virus genetic information assuming a DNA molecular weight of 108 and ... ranging in molecular weight from 15,000 to 280,000. Evidence for virus specificity based on increased rates of synthesis ... Identification and Relative Molar Rates of Synthesis of Structural and Nonstructural Herpes Virus Polypeptides in the Infected ...
The molecular weight of the CP was estimated by sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) on a 3.5 ... Protein molecular weight was estimated by comparing their relative mobility with molecular weight standards (Low molecular ... The predicted molecular weight of the protein is 30,563 Da, slightly larger than the 29,880 Da determined from the SDS-PAGE ( ... Amino acid analysis of the protein showed a molecular mass of 30,563 Da, close to the values obtained from electrophoresis ...
... sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels (Pierce, Rockford, IL) and then stained with SYPRO Ruby protein ... The lack of large molecular weight protein species in the elution of aged zebrafish lenses likely reflects the functional ... Relative abundance and/or percentage of total protein for gels from 10-day-, 6-week-, and 27-month-old samples were quantified ... and the resulting peaks analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE; 12.5% gel) to ...
... molecular weight, isoelectric point, relative content in microsomes, two-dimensional tryptic peptide maps, one-dimensional ... a single protein band on immunoblots of human liver microsomes after sodium dodecyl sulfate-polyacrylamide gel electrophoresis ...
More than 150 proteins could be identified by two-dimensional polyacrylamide gel electrophoresis. Proteins in the entire region ... from pH 7.0 to 9.0 with molecular weights (Mr) ranging from 35,000 to 41,000 and a protein focusing from pH 5.2 to 5.3 with Mr ... The electrophoretic patterns of nonhistone nuclear protein (NHNP) in MSS relative to N showed a striking resemblance to those ... demonstrated previously for the early stage (myolytic phase) of hamster cardiomyopathy relative to the matched control. Since ...
... molecular weight markers of the indicated sizes in kD. The relative molecular weight of the fused H chain of 75 kD in lane 1 (R ... The fusion protein was analyzed by electrophoresis on a 10% SDS-polyacrylamide gel (FIG. 8) under reducing (R) or non-reducing ... in gel sample buffer in the presence or absence of 2-mercaptoethanol and analyzed on a 7% polyacrylamide gel. Proteins were ... The position of stained marker proteins and their apparent molecular weights are indicated. The dried gel was exposed to film ...
On polyacrylamide gel electrophoresis, the circulating aggregate was indistinguishable from a material of similar molecular ... The relative roles of phospholipid fatty acyl chain length and phospholipid fatty acyl chain unsaturation in the determination ... 20 mg/kg body weight per wk) or diluent for 5, 10, and 15 wk. Animals were killed at these time periods and brush border ... When analyzed by polyacrylamide gel electrophoresis and Sepharose CL6B chromatography, the 440,000-D vWF oligomer is a dimer of ...
... polyacrylamide gel and electrotransferred from the slab gel to a nitrocellulose membrane. Prestained molecular weight markers, ... The PCR product was analyzed using gel electrophoresis and relative band intensity was determined. Results show the induction ... The PCR product was analyzed using gel electrophoresis and relative band intensity was determined. Results show the induction ... the major immunoreacted protein band within the appropriate molecular weight range was determined. The apparent molecular ...
SDS polyacrylamide gel and then electrophoresed with cooling. The molecular weight is determined relative to prestained ... methionine or cystein and polyacrylamide gel electrophoresis indicates that multiple molecular size forms of BMP-2A proteins ... In this gel system, the majority of bone and/or cartilage inductive fractions have the mobility of a protein having a molecular ... A. Molecular Weight. Approximately 20 ug protein from Example I is lyophilized and redissolved in 1X SDS sample buffer. After ...
... having an apparent molecular weight of less than about 15,000 daltons as determined by SDS-polyacrylamide gel electrophoresis ( ... 7, lanes 4 and 5, indicate that the 58 kDa form of huTF exhibits a higher relative molecular than the 47 kDa form because of ... Fifteen percent polyacrylamide gels were loaded as follows: lane 1 contains molecular weight standards with apparent molecular ... It was then subjected to SDS-polyacrylamide gel electrophoresis on a preparative-style slab gel as described by Laemmli, Nature ...
Polyacrylamide Gel Electrophoresis. The molecular weights of the glycosaminoglycan chains were estimated by polyacrylamide gel ... average molecular weight, 20 kD); St4, dextran sulfate (average molecular weight, 8 kD). D, Relative proportions of isomeric ... Gel filtration (Fig 7B⇓) and polyacrylamide gel electrophoresis (Fig 7C⇓) of this fraction confirmed its high molecular weight ... 6 we estimated the molecular weight of aortic glycosaminoglycans by gel filtration and polyacrylamide gel electrophoresis. The ...
... and high-molecular-weight markers (BioRad, Richmond, CA). Prior to electrophoresis, samples were adjusted to 2% (w/v) SDS and 3 ... SDS polyacrylamide gels were stained with Coomassie Blue R250 to visualize protein. Chemiluminescent Western blot analysis ... The relative abundance of the two proteins in these extracts was determined by comparison to purified samples. ... Molecular weight markers in kilobases are shown on the left side. (C) Autoradiogram of Southern blot of malA locus. Lane 1, ...
The molecular weight of the purified RoAmy determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis was ... The authors compared the risks by the age of 80 years for all cancers combined in female first-degree relatives of women with a ... Intra-molecular C-Hâ‹¯O hydrogen bonds occur. In the crystal, inter-molecular C-Hâ‹¯O hydrogen bonds link the mol-ecules into ... In present study, one nonpeptidyl small molecular weight compound (D34) was synthesized. Its SOD mimetic activity and the ...
Denaturing SDS vertical polyacrylamide gel electrophorCapillary electrophoresisVertical gel electrophoresis apparatusCharacterizationSodium dodecylMoleculesIsoelectric pointMigrateMigrationPolypeptidePolypeptidesDiscontinuous gelUtilizesBuffersChromatographyAcrylamide concentrationsQuantificationSubunitApparatusSlabAnalyticalLower acrylamideNucleic acidPreparativeSeparation of proteins basedConcentrationsBufferApparent molecular weightPurityCommonlyElectrophoretic runMicroheterogeneityMobilityPeptidePoreHigher molecular weight45,000AntibodyGradientDimensionalAnionicApproximate molecular weights35,000Separate proteinsMethodsSynthesisDaltonsGeneticsProteins basedProtein SeparationAutoradiographyProtocolsBiologyAmino acidsLaboratoryKnown molecularSubsequent
- Microchip based electrophoresis is a promising alternative to capillary electrophoresis since it has the potential to provide rapid protein analysis, straightforward integration with other microfluidic unit operations, whole channel detection, nitrocellulose films, smaller sample sizes and lower fabrication costs. (wikipedia.org)
- Adapun pemisahan dua tahap spesi Ca, tahap pertama dengan SEC dan tahap kedua dengan elektroforesis kapiler (capillary electrophoresis, CE). (itb.ac.id)
- Particularly, the fluorescent compositions find use as labels in analyzing double stranded and single stranded nucleic acid fragments using capillary electrophoresis. (rpxcorp.com)
- To find a suitable approach, three variants of RISA were tested: (1) standard, polyacrylamide gel-based, (2) automated, utilized capillary electrophoresis (GA-ARISA), and (3) automated microfluidics-based (MF-ARISA). (springer.com)
- High-Performance Capillary Electrophoresis Quantification of Global Histone H3 and H4 Acetylation. (pnas.org)
- This is the first report of the biological and molecular characterization of TSV isolated from soybeans. (scielo.br)
- Characterization of the immunoprecipitate of the mammary tissue carboxylase by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis reveals a single biotin-containing polypeptide of about 230000mol.wt. (biochemj.org)
- Molecular Characterization and Quantitative Analysis of Superoxide Dismutases in Virulent and Avirulent Strains of Aeromonas salmonicida subsp. (asm.org)
- Molecular characterization of the cyclic nucleotide-gated. (ubc.ca)
- 1989) of what is now known to be the CNG channel α subunit facilitated its molecular characterization. (ubc.ca)
- However, in the 1970, techniques developed for the recombinant DNA laboratory began to find application in the molecular characterization of clinical isolates. (elsevier.com)
- Sodium dodecyl sulphate polyacrylamide gel electrophoresis suggested the presence of three subunit types of molecular weights 56200, 53300 and 15700. (microbiologyresearch.org)
- SDS-PAGE Dr Anurag yadav,Bio-FMMC2 Sodium dodecyl sulphate- polyacrylamide gel electrophoresis. (sentinhell.fr)
- By using sodium dodecyl sulphate (SDS) and a gel made from acrylamide, protein shape, structure and charge no longer become factors as proteins migrate on to gels and protein bands are only affected by size. (sentinhell.fr)
- Silver ions are reduced to their metallic form by formaldehyde at alkaline pH.SDS-PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) is commonly used in the lab for the separation of proteins based on their molecular weight. (billsimas.com)
- The affinity purified lectin was found homogeneous by polyacrylamide gel electrophoresis, in sodium dodecyl sulphate-polyacrylamide gels, by gel filtration on Sephadex G-100 and by sedimentation velocity experiments. (iisc.ac.in)
- In sodium dodecyl sulphate-polyacrylamide gel electrophoresis the lectin gave a molecular weight of 24,000 in the presence as well as absence of 2-mercaptoethanol. (iisc.ac.in)
- The aim of this work was to illustrate benefits of performing a statistical molecular design (SMD) and proper statistical analysis of the molecules' properties before SAR and quantitative structure-activity relationship (QSAR) analysis. (diva-portal.org)
- The CD138/syndecan-1 isoform expressed by PEL has an average molecular weight of 420 kD, which is substantially different from that of CD138/syndecan-1 molecules generally expressed by plasma cells. (bloodjournal.org)
- The separation of molecules within a gel is determined by the relative size of the pores formed within the gel. (gac.edu)
- The higher the gel concentration, the smaller the pore size of the gel and the better it will be able to separate smaller molecules. (gac.edu)
- Tube gels have an advantage in that the movement of molecules through the gels is less prone to lateral movement and thus there is a slightly improved resolution of the bands, particularly for proteins. (gac.edu)
- Polyacrylamide gels restrain larger molecules from migrating as fast as smaller molecules. (rice.edu)
- The gradient is established before adding the particles of interest by first subjecting a solution of small molecules such as polyampholytes with varying pI values to electrophoresis. (wikipedia.org)
- When proteins are placed in an electric field, molecules with a net charge such as proteins, will move toward one electrode or the other, a phenomenon known as electrophoresis. (sentinhell.fr)
- In application to SDS-PAGE (random-coiled molecules and long-fiber gels), KR is proportional to molecular weight (MW). (sentinhell.fr)
- SDS Polyacrylamide Gel Electrophoresis (SDS-PAGE) is used to separate protein molecules based on size. (sentinhell.fr)
- Electrophoresis is the migration of charged molecules in solution in response to an electric field. (uct.ac.za)
- A porous gel may act as a sieve by retarding, or in some cases completely obstructing, the movement of large macromolecules while allowing smaller molecules to migrate freely. (uct.ac.za)
- Lower percentage gels are better for resolving very high molecular weight molecules, while much higher percentages of acrylamide are needed to resolve smaller proteins. (billsimas.com)
- Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is a method of separating molecules based on the difference of their molecular weight. (billsimas.com)
- Polyacrylamide gel separates protein molecules according to both size and charge. (bionity.com)
- In this gel, macro molecules separate according to their size. (bionity.com)
- Comparison of the EM-type cytochrome P-450 to that isolated from PM livers revealed no difference in regard to immuno-cross-reactivity, molecular weight, isoelectric point, relative content in microsomes, two-dimensional tryptic peptide maps, one-dimensional peptide maps with three proteases, amino acid composition, and amino-terminal protein sequence. (uniprot.org)
- Without SDS, different proteins with similar molecular weights would migrate differently due to differences in mass-charge ratio, as each protein has an isoelectric point and molecular weight particular to its primary structure. (billsimas.com)
- Glycosylated proteins may not migrate at their expected molecular weight because their migration is based more on the mass of their polypeptide chains, not the sugars that are attached (Sambrook J et al. (thermofisher.com)
- It must be noted that water cannot act as a substitute for one of these buffers, as the DNA will not migrate along the gel. (wikipedia.org)
- CRP was shown to migrate similarly to the two prolyl hydroxylase monomers which had molecular weights of 65,000 and 60,000. (uri.edu)
- Without SDS, different proteins with similar molecular weights would migrate differently due to differences in folding, as differences in folding patterns would cause some proteins to better fit through the gel matrix than others. (bionity.com)
- During electrophoresis in SDS-polyacrylamide gels there is a linear relationship between the logarithm of the molecular weight and the relative distance of the migration of the SDS-polypeptide micelle. (protocol-online.org)
- Determination of the molecular weight by relative electrophoretic mobilities on analytical polyacrylamide gels gave a higher value, 225,000, thus demonstrating that the 9s RNA had different migration properties compared to the rRNA 174 marker species. (gla.ac.uk)
- In a gel of uniform density the relative migration distance of a protein (Rf, the f as a subscript) is negatively proportional to the log of its mass. (rice.edu)
- Gels with large pores are usually used in this process to eliminate any "sieving" effects, or artifacts in the pI caused by differing migration rates for proteins of differing sizes. (wikipedia.org)
- A molecular-weight size marker, also referred to as a protein ladder, DNA ladder, or RNA ladder, is a set of standards that are used to identify the approximate size of a molecule run on a gel during electrophoresis, using the principle that molecular weight is inversely proportional to migration rate through a gel matrix. (wikipedia.org)
- The gel percentage effects the migration of the DNA. (wikipedia.org)
- The molecular weight is determined by comparing the migration of protein spots to the migration of standards. (sentinhell.fr)
- In denaturing SDS-PAGE separations therefore, migration is determined not by intrinsic electrical charge of the polypeptide, but by molecular weight . (uct.ac.za)
- SDS-PAGE separates proteins according to their molecular weight, based on their differential rates of migration through a sieving matrix a gel under the influence of an applied electrical field. (billsimas.com)
- The SDS binds to the protein in a ratio of approximately 1.4 g SDS per 1.0 g protein (although binding ratios can vary from 1.1-2.2 g SDS/g protein), giving an approximately uniform mass:charge ratio for most proteins, so that the distance of migration through the gel can be assumed to be directly related to only the size of the protein. (bionity.com)
- Construction and expression of a truncated form of the (3 subunit demonstrated that it is the glutamic acidrich N-terminal portion of the (3 subunit that is responsible for its anomalous migration on an SDS gel. (ubc.ca)
- A polypeptide chain binds amounts of SDS in proportion to its relative molecuar mass. (rice.edu)
- SDS PAGE separates proteins according to their electrophoretic mobility, reflecting their molecular weight (size of polypeptide). (nabas.no)
- A linear relationship exists between the logarithm of the molecular weight of an SDS-denatured polypeptide, or native nucleic acid, and its Rf . (uct.ac.za)
- SDS-PAGE , officially sodium dodecyl sulfate polyacrylamide gel electrophoresis , is a technique used in biochemistry , genetics and molecular biology to separate proteins according to their electrophoretic mobility (a function of length of polypeptide chain or molecular weight as well as higher order protein folding , posttranslational modifications and other factors). (bionity.com)
- Analyses of polypeptides made in HEp-2 cells infected with herpes simplex virus type 1 by high-resolution polyacrylamide gel electrophoresis revealed the synthesis of at least 49 infected cell polypeptides (ICP) ranging in molecular weight from 15,000 to 280,000. (asm.org)
- These 47 polypeptides can account for 75% of the virus genetic information assuming a DNA molecular weight of 10 8 and asymmetric transcription. (asm.org)
- On the basis of their mobility relative to virion proteins, the ICP were classified as structural (S, 23 polypeptides), nonstructural (NS, 16 polypeptides), and unassigned (U, 10 polypeptides). (asm.org)
- Conversely, gels up to 30% have been used to separate small polypeptides. (gac.edu)
- Because the charge-to-mass ratio is nearly the same among SDS-denatured polypeptides, the final separation of proteins is dependent almost entirely on the differences in relative molecular mass of polypeptides. (rice.edu)
- The lower layer (separating, or resolving, gel) is responsible for actually separating polypeptides by size. (rice.edu)
- Analysis of the major protein components of the membranes of fractionated cells by SDS-polyacrylamide gel electrophoresis indicated that aging was accompanied by the appearance of two new polypeptides of approximate molecular weights 63,000 and 25,000, with probable degradation of component 3- an extracellular surface membrane glycoprotein. (gla.ac.uk)
- Analysis of purified Oriboca virions by neutral, sodium dodecyl sulfate polyacrylamide-gel electrophoresis indicated the presence of three structural polypeptides designated V-1, V-2, and V-3 on the basis of their relative electrophoretic mobilities in 8% gels. (asm.org)
- Molecular weights have been tentatively assigned to the polypeptides by extrapolation from the structural polypeptides of Sindbis virus when both are run in the same gel. (asm.org)
- This material is accompanied by a presentation on protein structure and principles behind denaturing samples and discontinuous gel electrophoresis. (rice.edu)
- During electrophoresis in a discontinuous gel system, an ion gradient is formed in the early stage of electrophoresis that causes all of the proteins to focus into a single sharp band. (billsimas.com)
- Reference to the Laemmli method in a materials and methods section eliminates the need to describe the buffers, casting of gels, apparatus, etc. (rice.edu)
- This classic system uses a discontinuous buffer system where the pH and ionic strength of the buffer used for running the gel (Tris, pH 8.3) is different from the buffers used in the stacking gel (Tris, pH 6.8) and resolving gel (Tris, pH 8.8). (thermofisher.com)
- In DNA electrophoresis, the TAE (Tris-acetate-EDTA) and TBE (Tris-borate-EDTA) are the usual buffers of choice. (wikipedia.org)
- Columns and matrix, GFP and BFP extracts protein, buffers and reagents for electrophoresis. (bioted.es)
- In the second dimension, protein separation is performed based on molecular weight using SDS Laemmli or Tris-Tricine buffers. (biomedcentral.com)
- Two-dimensional gel electrophoresis and size exclusion chromatography were used to characterize the lens crystallin content of 4.5-day to 27-month-old zebrafish. (molvis.org)
- Constituents of size exclusion chromatography elution peaks were identified with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. (molvis.org)
- SDS-Denaturing Protein Electrophoresis (Electrophoresis Package 2 & 2M - Page ) Molecular Weight Determination 14/16 Human and Bacterial Amylase 14 Peptide Mapping Analysis 14/17 Protein Evolution and the Western Blot 14/17 Affinity Chromatography 15/17 Contractile Proteins from Cow Heart 16 Isolation of Chromosomal Proteins 16/17. (sentinhell.fr)
- Metode pemisahan utama dilakukan dengan menggunakan kromatografi eksklusi ukuran (size exclusion chromatography, SEC) sedangkan metode pemisahan pendukung adalah quantitative preparative native continuous polyacrylamide gel electrophoresis (QPNC PAGE). (itb.ac.id)
- The relative molecular weight of this lectin is determined to be 48,000 ± 1,000 by gel chromatography and sedimentation equilibrium experiments. (iisc.ac.in)
- employing diethyl aminoethyl cellulose ion exchange and Biogel P-150 gel filtration column chromatography. (springeropen.com)
- Hydrophobic interaction chromatography Molecular size 1. (powershow.com)
- Gel filtration chromatography 4. (powershow.com)
- These data provide the first two-dimensional gel electrophoresis maps of the developing zebrafish lens, with quantification of changing crystallin abundance and visualization of post-translational modification. (molvis.org)
- Isobaric tags for relative and absolute quantification (iTRAQ) were used to identify urinary proteins that were differentially excreted in normoalbuminuric and microalbuminuric patients with type 2 diabetes where 710 and 196 proteins were identified and quantified, respectively. (hindawi.com)
- In combination with Quantity One and PD Quest TM softwares used for relative protein quantification. (mpg.de)
- Analytical sodium dodecyl sulfate-polyacrylamide gel electrophoresis of human spinal cord components followed by immunoblots with sera under study revealed that the serum antibody was specific for the high molecular weight protein subunit of neurofilaments. (jci.org)
- The cloned CNG channel β subunit cDNA codes for a protein with a predicted molecular mass of 155 kDa (Korschen et al, 1995). (ubc.ca)
- Figure 4.1 shows a typical slab electrophoresis unit. (gac.edu)
- One, column electrophoresis, uses tubular gels formed in glass tubes, while the other, slab gel electrophoresis, uses flat gels formed between two plates of glass. (gac.edu)
- CRP was purified by antibody precipitation from the dialyzed 70% (NH4)2S04 supernatants and subsequent electrophoresis on 10% SDS polyacrylamide slab gels. (uri.edu)
- One dimensional electrophoresis is used for most routine protein and nucleic acid separations. (gac.edu)
- Nucleic acids however, remain negative at any pH used for electrophoresis and in addition carry a fixed negative charge per unit length of molecule, provided by the PO4 group of each nucleotide of the the nucleic acid. (uct.ac.za)
- The technique of preparative polyacrylamide gel electrophoresis was adapted for the isolation of pure 9n RNA from total 14 day mouse embryo liver nucleic acids. (gla.ac.uk)
- Preliminary experiments used the M.S.E. B-XV zonal rotor for the isolation of a fraction enriched in the 148 mIINP particle dissociated from polysomes by EBT.A. The RNA isolated from the 14s mRNP particle was further purified by preparative gel electrophoresis. (gla.ac.uk)
- For example, quantitative preparative native continuous polyacrylamide gel electrophoresis QPNC-PAGE is a method for separating native metalloproteins in complex biological matrices. (billsimas.com)
- The low levels of zebrafish lens α-crystallin relative to mammals may be due to the high concentrations of γ-crystallins in this aquatic lens. (molvis.org)
- Glucan synthesis by free or glucan-bound glucosyltransferase was stimulated by low concentrations (1 to 5 mg ml −1 ) of isomaltose or water-soluble dextrans of various molecular weights, but higher concentrations (10 mg ml −1 ) inhibited glucan synthesis. (microbiologyresearch.org)
- Polyacrylamide, which is easy to handle and to make at higher concentrations, is used to separate most proteins and small oligonucleotides that require a small gel pore size for retardation. (uct.ac.za)
- As the name suggests, the gel matrix used for SDS-PAGE is polyacrylamide, which is a good choice because it is chemically inert and, crucially, can easily be made up at a variety concentrations to produce different pore sizes giving a variety of separating conditions that can be changed depending on your needs. (billsimas.com)
- The extracted protein must be dissolved and preserved in lysis buffer to further conduct the following electrophoresis process. (creative-diagnostics.com)
- In electrophoresis, proteins are separated on the basis of charge, and the charge of a protein can be either + or -- , depending upon the pH of the buffer. (gac.edu)
- Factors such as buffer, charge/voltage, and concentration of gel can affect the mobility and/or appearance of your marker/ladder/standard. (wikipedia.org)
- Furthermore, using water instead of buffer will result in the gel melting. (wikipedia.org)
- There are two types of buffer systems in electrophoresis, continuous and discontinuous . (uct.ac.za)
- Kondisi operasional yang diterapkan meliputi (i) sistem buffer kontinyu 20 mM Tris-HCl/1 mM NaN3 pH 10, (ii) derajat polimerisasi poliakrilamid 4 %T / 2,67 % C, panjang gel 40 mm, fasa gerak 20 mM MES/1 mM NaN3 pH 8,0, dan (iii) temperatur analisis 4 derajat C. Sampel difraksinasi sebanyak 74 fraksi dengan volume setiap fraksi 5 mL (fraksinasi otomatis dengan pemograman). (itb.ac.id)
- To ensure that the sample sinks to the bottom of the gel, sample buffer is supplemented with additives that increase the density of the sample. (billsimas.com)
- This gel is prepared with Tris buffer pH 6.8 of about 2 pH units lower than that of electrophoresis buffer. (bionity.com)
- The most commonly used technique is sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). (thermofisher.com)
- Polyacrylamide gel electrophoresis (SDS-PAGE) Gel electrophoresis of proteins with a polyacrylamide matrix, commonly called polyacrylamide gel electrophoresis (PAGE) is undoubtedly one of the most widely used techniques to characterize complex protein mixt ures. (sentinhell.fr)
- The most widely used gel system for separating a broad range of proteins by SDS-PAGE is the Laemmli system (Nature 227:680 (1970)), which uses Tris-glycine gels comprising a stacking gel component that helps focus the proteins into sharp bands at the beginning of the electrophoretic run and the resolving gel, where higher gel percentages separate the proteins based on their size. (thermofisher.com)
- Protein molecular weight markers are used to calculate sample molecular weights, to monitor the progress of an electrophoretic run, or as a positive control for analysis conditions. (sentinhell.fr)
- A tracking dye may be added to the protein solution to allow the experimenter to track the progress of the protein solution through the gel during the electrophoretic run. (bionity.com)
- Protein electrophoresis is the process of separating proteins by placing them in a gel matrix and then observing protein mobility in the presence of an electrical field. (thermofisher.com)
- In these gels, protein mobility is a function of the protein's length and charge. (thermofisher.com)
- Prestained Proteins with unknown molecular weights are assigned molecular weights based on the relative mobility of prestained standard protein markers. (skolebutik.dk)
- The isolates showed 3 LPS patterns differing in the number and electrical mobility of bands in silver-stained gel. (bvsalud.org)
- The pore size of a gel is determined by two factors, the total amount of acrylamide present (designated as %T) and the amount of cross-linker (%C). As the total amount of acrylamide increases, the pore size decreases. (gac.edu)
- It is a synthetic gel, thermo-stable, transparent, strong, relatively chemically inert, can be prepared with a wide range of average pore sizes, can withstand high voltage gradients, feasible to various staining and destaining procedures and can be digested to extract separated fractions or dried for autoradiography and permanent recording. (bionity.com)
- whereas preincubation with higher molecular weight dextrans significantly inhibited the glucosyltransferase binding. (microbiologyresearch.org)
- However, only the higher molecular weight components were used in the turnover studies of CRP. (uri.edu)
- In general, proteins with the greatest specific radioactivities are those in the higher molecular weight range. (springer.com)
- When the reaction mixture contained EGTA and no added Ca 2+ , 32 P was incorporated into two proteins with molecular weights of 45,000 and 13,000. (diabetesjournals.org)
- Use 5% gels for proteins ranging from 60,000 to 200,000 daltons, 10% gels for a range of 16,000 to 70,000 daltons and 15% gels for a range of 12,000 to 45,000 daltons. (gac.edu)
- Antibody is incorporated into liquefied agar and allowed to gel. (skolebutik.dk)
- The antigen is added to small wells and radiates throughout the antibody-containing medium, leaving a precipitate throughout the gel. (skolebutik.dk)
- Test results from Biacore measurements indicate binding capacity of the antibody by comparison to curve with relative number describing binding capacity. (nabas.no)
- HDL particle size by gradient gel electrophoresis revealed small HDL particles (estimated Stokes' diameter, 8.14 to 8.30 nm). (ahajournals.org)
- IEF involves adding an ampholyte solution into immobilized pH gradient (IPG) gels. (wikipedia.org)
- IPGs are the acrylamide gel matrix co-polymerized with the pH gradient, which result in completely stable gradients except the most alkaline (>12) pH values. (wikipedia.org)
- Among them, Denaturing Gradient Gel Electrophoresis (DGGE) analysis is one of the most widely-used molecular techniques, enabling the identification of community members by the recovery and sequencing of amplification products. (springer.com)
- Separation of proteins under a pH gradient allows intense band recovering using various tactics such as immobilized gradient electrophoresis (IPEG), isoelectric focusing (IEF), or non-equilibrium pH gradient electrophoresis (NEPHGE). (biomedcentral.com)
- More than 150 proteins could be identified by two-dimensional polyacrylamide gel electrophoresis. (ahajournals.org)
- Electrophoresis can be one dimensional (i.e. one plane of separation) or two dimensional. (gac.edu)
- The gels are neutral, hydrophillic, three-dimensional networks of long hydrocarbons crosslinked by methylene groups. (gac.edu)
- Isoelectric focusing is the first step in two-dimensional gel electrophoresis, in which proteins are first separated by their pI and then further separated by molecular weight through SDS-PAGE. (wikipedia.org)
- Two-dimensional gel electrophoresis is an invaluable tool that provides insights into protein complexes and sub-organelle organization. (billsimas.com)
- Among these methods, two dimensional- polyacrylamide gel electrophoresis (2-DE) represents a mainstay orthogonal approach, which is popularly used to simultaneously fractionate, identify, and quantify proteins when coupled with mass spectrometric identification or other immunological tests. (biomedcentral.com)
- Two dimensional polyacrylamide gel electrophoresis (2-DE) is considered a powerful tool used for separation and fractionation of complex protein mixtures from tissues, cells, or other biological samples. (biomedcentral.com)
- Proteins in the entire region from pH 7.0 to 9.0 with molecular weights (Mr) ranging from 35,000 to 41,000 and a protein focusing from pH 5.2 to 5.3 with Mr of 55,000 were strikingly reduced in MSS. (ahajournals.org)
- The protein had slightly different molecular masses in the Mr range of 35,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. (nih.gov)
- Novex® precast protein gels: the beauty of a straight lineAs discussed earlier, the key use of protein gels is to separate proteins for subsequent transfer onto a membrane for interrogation with antibodies, a process called western blotting. (thermofisher.com)
- Protein gel electrophoresis is a simple way to separate proteins prior to downstream detection. (sentinhell.fr)
- SDS is used to create denaturing conditions to separate proteins by molecular weight and also confers negative charge to the proteins in proportion to its mass. (sentinhell.fr)
- Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is used to separate proteins with relative molecular mass no smaller than 10 KD. (sentinhell.fr)
- To separate proteins in an electrical field based on their molecular weight only, we need to destroy the tertiary structure by reducing the protein to a linear molecule, and somehow mask the intrinsic net charge of the protein. (billsimas.com)
- Unless the paper employs some modification to the method, the only details of SDS-PAGE that should be reported in a methods section are percent total acrylamide (%T) in a gel, relative percentage and type of crosslinker (%C), and perhaps a reference to the gel dimensions. (rice.edu)
- There are two common methods in which to construct a DNA molecular-weight size marker. (wikipedia.org)
- Recently, a study of complex microbial communities has been performed by the application of culture-independent, molecular methods based on directed analysis of the 16S rRNA gene structure. (springer.com)
- Gel- based proteomics is one of the most versatile methods for fractionating protein complexes. (biomedcentral.com)
- Molecular mass (symbol m) is expressed in Daltons (Da). (rice.edu)
- It is incorrect to express molecular weight (relative molecular mass) in Daltons. (rice.edu)
- Nevertheless you will find the term molecular weight used with Daltons or kiloDaltons in some literature, often using the abbreviation MW for molecular weight. (rice.edu)
- The 32 P-labeled proteins were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the 32 P content was visualized by autoradiography, and the relative intensities of specific bands were quantitated. (diabetesjournals.org)
- The sodA gene encoded a protein of 204 amino acids with a molecular mass of approximately 23.0 kDa (SodA) that had high similarity to other prokaryotic Mn-SODs. (asm.org)
- And it has a molecular weight of 16700 Da and 153 amino acids (4). (ukessays.com)
- Adding SDS solves this problem, as it linearizes the proteins so that they may be separated strictly by molecular weight ( primary structure , or number (and size) of amino acids ). (bionity.com)