电泳工作原理（Electrophoresis principle）.docVIP

电泳工作原理（Electrophoresis principle） Working principle Protein separation is one of the important biological chemical separation and purification technology, electrophoresis refers to charged particles in the electric field, and toward the opposite charge electrode movement phenomenon. According to the support of the different, with agarose gel electrophoresis, starch gel electrophoresis and polyacrylamide gel electrophoresis. The polyacrylamide gel electrophoresis (PAGE) because there is no electroosmosis, small amount of sample (1-100 g), high resolution, can be detected in 10-9-10-12mol gel samples, high mechanical strength, good repeatability and can be adjusted by the monomer concentration or the ratio of monomer and crosslinking agent and the advantages of different aperture gel has been widely used. SDSis the most common method of qualitative analysis of protein electrophoresis, especially for protein purity detection and determination of protein molecular weight PAGE can separate proteins effectively, mainly based on the difference between the molecular weight and charge, and SDS(SDS polyacrylamide gel electrophoresis) the separation principle is only based on the difference of protein molecular weight, because the liquid sample of SDSis to run in electrophoresis sample if containing SDS and mercaptoethanol (2-ME) two thiol or erythritol (DTT), which can disconnect the two disulfide bonds between cysteine residues, four stage structural damage proteins, SDS is a kind of anionic surfactant or detergent, it can disconnect the intramolecular and intermolecular hydrogen bonds, destroy protein two level and three level structure, and combined with the hydrophobic part of the protein, the destruction of their folding structure, if the electrophoresis sample sample buffer, to cook 3-5 minutes to fully combine to form the SDS- SDS and protein protein complexes in the boiling water, SDS- protein complexes In strong reducing agent in the presence of mercaptoethanol, two