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Protein purification

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Protein purification is a series of processes intended to isolate one or a few proteins from a complex mixture, usually cells, tissues, or whole organisms. Protein purification is vital for the specification of the function, structure, and interactions of the protein of interest. The purification process may separate the protein and non-protein parts of the mixture, and finally separate the desired protein from all other proteins.

ChromatographyChromatographySet of laboratory techniques for separation of mixtures Thin-layer chromatography is used to separate components of a plant extract, illustrating the experiment with plant pigments which gave chromatography its name In chemical analysis, chromatography is a laboratory technique for the separation of a mixture into its components.SDS-PAGESDS-PAGESDS-PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis) is a discontinuous electrophoretic system developed by Ulrich K. Laemmli which is commonly used as a method to separate proteins with molecular masses between 5 and 250 kDa. The combined use of sodium dodecyl sulfate (SDS, also known as sodium lauryl sulfate) and polyacrylamide gel eliminates the influence of structure and charge, and proteins are separated by differences in their size.Size-exclusion chromatographySize-exclusion chromatographySize-exclusion chromatography, also known as molecular sieve chromatography, is a chromatographic method in which molecules in solution are separated by their shape, and in some cases size. It is usually applied to large molecules or macromolecular complexes such as proteins and industrial polymers.Denaturation (biochemistry)Denaturation (biochemistry)IUPAC definitionProcess of partial or total alteration of the native secondary, and/or tertiary, and/or quaternary structures of proteins or nucleic acids resulting in a loss of bioactivity. Note 1: Modified from the definition given in ref. Note 2: Denaturation can occur when proteins and nucleic acids are subjected to elevated temperature or to extremes of pH, or to nonphysiological concentrations of salt, organic solvents, urea, or other chemical agents.Sodium dodecyl sulfateSodium dodecyl sulfateSodium dodecyl sulfate (SDS) or sodium lauryl sulfate (SLS), sometimes written sodium laurilsulfate, is an organic compound with the formulaCH3(CH2)11OSO3Na and structure H3C−(CH2)11−O−S(=O)2−ONa. It is an anionicsurfactant used in many cleaning and hygiene products. This compound is the sodium salt of the 12-carbon organosulfate. Its hydrocarbon tail combined with a polar "headgroup" give the compound amphiphilic properties that make it useful as a detergent.ProteaseProteaseA protease (also called a peptidase, proteinase, or proteolytic enzyme) is an enzyme that catalyzesproteolysis, breaking down proteins into smaller polypeptides or single amino acids, and spurring the formation of new protein products. They do this by cleaving the peptide bonds within proteins by hydrolysis, a reaction where water breaks bonds.AntibodyAntibodyAn antibody (Ab), or immunoglobulin (Ig), is a large protein belonging to the immunoglobulin superfamily which is used by the immune system to identify and neutralize antigens such as those that exist on bacteria and virus cells, including those that cause disease. Each individual antibody recognizes one or more specific antigens, and antigens (a portmanteau of "antibody generator") of virtually any size and chemical composition can be recognized.ProteinProteinBiomolecule consisting of chains of amino acid residuesA representation of the 3D structure of the protein myoglobin showing turquoise α-helices. This protein was the first to have its structure solved by X-ray crystallography. Toward the right-center among the coils, a prosthetic group called a heme group (shown in gray) with a bound oxygen molecule (red).Proteins are large biomolecules and macromolecules that comprise one or more long chains of amino acidresidues.Triton X-100Triton X-100Triton X-100 (C14H22O(C2H4O)n) is a nonionic surfactant that has a hydrophilic polyethylene oxide chain (on average it has 9.5 ethylene oxide units) and an aromatic hydrocarbon lipophilic or hydrophobic group. The hydrocarbon group is a 4-(1,1,3,3-tetramethylbutyl)-phenyl group. Triton X-100 is closely related to IGEPAL CA-630, which might differ from it mainly in having slightly shorter ethylene oxide chains.Biological activityIn pharmacology, biological activity or pharmacological activity describes the beneficial or adverse effects of a drug on living matter. When a drug is a complex chemical mixture, this activity is exerted by the substance's active ingredient or pharmacophore but can be modified by the other constituents.Cell biologyCell biologyCell biology, cellular biology, or cytology, is the branch of biology that studies the structure, function, and behavior of the cells. All organisms are made of cells. A cell is the basic unit of life that is responsible for the living and functioning of an organism. Cell biology encompasses both prokaryotic and eukaryotic cells, with subtopics including the study of cell metabolism, cell communication, cell cycle, biochemistry, and cell composition.Tissue (biology)Tissue (biology)In biology, tissue is an assembly of similar cells and their extracellular matrix from the same embryonic origin that together carry out a specific function. Tissues occupy a biological organizational level between cells and a complete organ. Accordingly, organs are formed by the functional grouping of multiple tissues. The English word "tissue" derives from the French word "", meaning "fabric", derived from the verb “tisser”, "to weave".

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