sds-page电泳过程中常见问题以及解决方法（Common problems and solutions in the process of SDS-PAGE electrophoresis）.docVIP

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sds-page电泳过程中常见问题以及解决方法（Common problems and solutions in the process of SDS-PAGE electrophoresis）.doc

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sds-page电泳过程中常见问题以及解决方法（Common problems and solutions in the process of SDS-PAGE electrophoresis）

sds电泳过程中常见问题以及解决方法（Common problems and solutions in the process of SDSelectrophoresis） Common problems and solutions in the process of SDS Almost all the protein electrophoresis analysis was carried out in polyacrylamide gel, and all conditions to ensure that the total protein dissociated into single polypeptide subunits and may reduce their mutual aggregation, is the most commonly used SDS PAGE electrophoresis on the frequently encountered problems in the process we are discussed: The basic principle of Q:SDSPAGE electrophoresis? A:SDS- polyacrylamide gel electrophoresis in polyacrylamide gel, is the introduction of SDS system (twelve sodium dodecyl sulfate), SDS peptide and protein denaturation, and the negative charge, the peptide binding amount of SDS is almost always associated with the molecular weight of peptides and its sequence is proportional to the independent, so SDS peptide complex migration in polyacrylamide gel the rate of electrophoresis is only related to the size of a polypeptide, in saturated state, per gram of peptide binding to 1.4g detergent. When the molecular weight from 15KD to 200KD, there is a linear relationship between the logarithm of the migration rate and the molecular weight of protein, in accordance with the following formula: logMW=K-bX, where MW is the molecular weight of X, K, B migration rates are constant, if the migration of standard proteins of known molecular weight ratio on molecular weight of the number of mapping, can obtain a standard curve, unknown protein electrophoresis under the same conditions, according to its electrophoretic mobility can be obtained on the standard curve of molecular weight. Q: the effect of gel buffer system on electrophoresis? A: in SDS PAGE discontinuous electrophoresis, the gel buffer uses the Tris HCL buffer system, the concentrated gel is pH6.7, the separation gel pH8.9; and the electrophoretic buffer uses the Tris glycine buffer system. In concentrated gum, the pH environment is weak acidit