Native polyacrylamide gels.
Native polyacrylamide gels.
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DOI：
10.1007/978-1-61779-821-4_5
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发表时间：
2012-01-01
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影响因子：
--
通讯作者：
Bachmann, Michael
中科院分区：
文献类型：
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作者：
Arndt, Claudia;Koristka, Stefanie;Bachmann, Michael
通常，蛋白质在去污剂存在下，在（热）变性和（非）还原条件下通过聚丙烯酰胺凝胶电泳（PAGE）分离。最常用的洗涤剂是十二烷基硫酸钠（SDS）。SDS的主要功能是屏蔽待分析混合物中存在的蛋白质的相应电荷，并为所有蛋白质提供负电荷。因此，蛋白质将根据其分子量分离。蛋白质的电泳也可以在不存在SDS的情况下进行。使用这样的“天然”条件，每种蛋白质的电荷将主要影响电泳期间相应蛋白质的迁移率，所述电荷将取决于蛋白质的一级氨基酸序列（等电点）和电泳期间的pH。在这里，我们描述了一个“天然”PAGE的起始协议。
Usually proteins are separated by polyacrylamide gel electrophoresis (PAGE) in the presence of a detergent and under (heat-) denaturing and (non- or) reducing conditions. The most commonly used detergent is sodium dodecyl sulfate (SDS). The major function of SDS is to shield the respective charge of the proteins present in the mixture to be analyzed and to provide all proteins with a negative charge. As a consequence, the proteins will be separated according to their molecular weight. Electrophoresis of proteins can also be performed in the absence of SDS. Using such "native" conditions, the charge of each of the proteins, which will depend on the primary amino acid sequence of the protein (isoelectric point) and the pH during electrophoresis, will mainly influence the mobility of the respective protein during electrophoresis. Here we describe a starting protocol for "native" PAGE.