List, discuss, and evaluate the major technique used in separating proteins, including: solubility, size, dialysis, SDS-PAGE, charge-ion...

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Biology

List, discuss, and evaluate the major technique used inseparating proteins, including: solubility, size, dialysis,SDS-PAGE, charge-ion exchange chromatography, size and charge 2Dgels, specific ligand binding affinity chromatography,antigen-antibody recognition (western blot, ELISA); massspectrometry of peptide and proteomics

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Answer Purpose of separation The separation or purification step is essential to separate protein and non protein parts of mixtures The following techniques used to separate the desired protein from the mixture Based on Size Based on the size the proteins are used to separate from a mixture by using porous gels The basic principle is that smaller molecules have to transverse a larger volume in a porous matrix Based on the solubility Proteins are precipitated out from aqueous solutions when added salt concentration exceeds a critical level is known as salting out Because the salts bound to the water and there is no water to hydrate the proteins 1 Salting out process The separation of the proteins based o solubility occurs in two steps 1 Salt is added at a concentration just below necessary to precipitate out the protein of interest The solution is centrifuged to remove if any See Answer

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