Native Polyacrylamide Gels
Native Polyacrylamide Gels
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DOI：
10.1007/978-1-4939-8793-1_8
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发表时间：
2019-01-01
期刊：
影响因子：
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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
在有洗涤剂存在的条件下，在(热)变性和(或)还原条件下，用聚丙烯酰胺凝胶电泳法(PAGE)可以很容易地分离蛋白质。最常用的洗涤剂是十二烷基硫酸钠(十二烷基硫酸钠)。十二烷基硫酸钠的主要功能是屏蔽待分析混合物中蛋白质各自的电荷，并为所有蛋白质提供负电荷。因此，这些蛋白质将根据它们的分子量进行分离。蛋白质的电泳法也可以在没有十二烷基硫酸钠的情况下进行。在这种“天然”条件下，每种蛋白质的电荷将取决于蛋白质的初级氨基酸序列(等电点)和电泳过程中的pH值，这将主要影响各自蛋白质在电泳过程中的迁移率。这里我们描述了一个用于“原生”页面的启动协议。
Proteins can easily be 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.