sds-page电泳常见问题的分析（Analysis of common problems of SDS-PAGE electrophoresis）.docVIP

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sds-page电泳常见问题的分析（Analysis of common problems of SDS-PAGE electrophoresis）.doc

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sds-page电泳常见问题的分析（Analysis of common problems of SDS-PAGE electrophoresis）

sds电泳常见问题的分析（Analysis of common problems of SDSelectrophoresis） 1. effect of gel buffer system on electrophoresis? In SDS PAGE discontinuous electrophoresis, the gel buffer was used in the Tris HCL buffer system, the concentrated gel was pH6.7, and the separation gel pH8.9; while the electrophoretic buffer used the Tris glycine buffer system. In concentrated gum, the pH environment is weak acidity, so the glycine dissociation is very few, under the effect of the field, swimming efficiency is low; and the CL ion is very high, the formation of lower conductivity zone between the two, between protein molecules between the two in swimming. Due to the conductivity and electric field intensity is inversely proportional to the belt formed in the area of high voltage gradient, pressure of protein molecules together, condensed into a narrow zone. When the sample into the separating gel, due to the increase in pH, glue is alkaline, a large number of glycine dissociation rate increased after swimming, directly followed the chloride ion, at the same time as the separation gel pore size reduced, under the action of electric field, the protein molecules were separated according to charge and its inherent molecular size. Therefore, the effect of pH on the whole reaction system is crucial. In the experiment, after the exclusion of other factors, it is still not a good solution to the problem, and the factor should be considered first. 2. how are the samples treated? According to the different purposes of sample separation, there are three main treatment methods: reduced SDS treatment, non reduction SDS treatment, and reductive SDS treatment with alkylation. 1) reduction of SDS and DTT in the treatment: adding SDS sample in buffer (or Beta mercaptoethanol), protein conformation by dissociation, charge is neutralized, the formation of molecular SDS and protein combination, in electrophoresis, only separated according to their molecular weight. General electrophoresis is processed in