Sasse Joachim, Gallagher Sean R
Shriners Hospital for Crippled Children, Tampa, Florida, USA.
Curr Protoc Immunol. 2004 Feb;Chapter 8:Unit 8.9. doi: 10.1002/0471142735.im0809s58.
This unit describes protocols for detecting protein in a gel by either Coomassie blue, silver or fluorescent staining. Alternate rapid staining procedures are provided for each method and a support protocol describes how to photograph stained gels. Fluorescent staining (e.g., with SYPRO Orange or Red) is described as a popular alternative to traditional staining procedures, mainly because it is more sensitive than Coomassie staining, and often as sensitive as silver staining. Staining of proteins in SDS-polyacrylamide gels is described, and an alternate protocol details variations in the procedure for proteins in nondenaturing gels. A final support protocol describes the photography of fluorescently stained proteins.
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。每种方法都提供了替代的快速染色程序,一个辅助方案描述了如何拍摄染色后的凝胶。荧光染色(如使用SYPRO Orange或Red)被描述为传统染色程序的一种流行替代方法,主要是因为它比考马斯染色更灵敏,且通常与银染一样灵敏。文中描述了SDS聚丙烯酰胺凝胶中蛋白质的染色方法,另一个替代方案详细说明了非变性凝胶中蛋白质染色程序的变化。最后一个辅助方案描述了荧光染色蛋白质的拍摄方法。