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对凝胶中的蛋白质进行染色。

Staining proteins in gels.

作者信息

Sasse Joachim, Gallagher Sean R

机构信息

Shriners Hospital for Crippled Children, Tampa, Florida, USA.

出版信息

Curr Protoc Mol Biol. 2009 Jan;Chapter 10:Unit 10.6. doi: 10.1002/0471142727.mb1006s85.

Abstract

This unit describes protocols for detecting protein in a gel by Coomassie blue, silver, or fluorescent staining. As a general protein stain, Coomassie is easier and more rapid; however, fluorescent and silver staining methods are considerably more sensitive and thus can be used to detect smaller amounts of protein. Fluorescent staining is a popular alternative to traditional staining procedures, mainly because it is more sensitive than Coomassie staining, and often as sensitive as silver staining. Alternate protocols describe rapid Coomassie and silver staining methods, as well as fluorescent stains that are specific for phosphoproteins and glycoproteins. Staining of proteins in SDS-polyacrylamide gels is described; variations for fluorescent staining of proteins in nondenaturing gels are also included. Support protocols describe photography of stained proteins.

摘要

本单元介绍了通过考马斯亮蓝、银染或荧光染色检测凝胶中蛋白质的方法。作为一种通用的蛋白质染色方法,考马斯亮蓝染色操作更简便、速度更快;然而,荧光染色和银染方法灵敏度要高得多,因此可用于检测含量更少的蛋白质。荧光染色是传统染色方法的一种流行替代方法,主要是因为它比考马斯亮蓝染色更灵敏,且通常与银染灵敏度相当。其他方法介绍了快速考马斯亮蓝和银染方法,以及针对磷酸化蛋白和糖蛋白的荧光染料。文中描述了SDS聚丙烯酰胺凝胶中蛋白质的染色方法;还包括非变性凝胶中蛋白质荧光染色的变体。辅助方法介绍了染色蛋白质的拍照方法。

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