Sasse Joachim, Gallagher Sean R
Shriners Hospital for Crippled Children, Tampa, Florida, USA.
Curr Protoc Mol Biol. 2003 Nov;Chapter 10:Unit10.7. doi: 10.1002/0471142727.mb1007s64.
In the basic and alternate protocols of this unit, proteins are stained after electroblotting from polyacrylamide gels to blot transfer membranes. If the samples of interest are electrophoresed in duplicate and transferred to a blot transfer membrane, half of the membrane can be stained to determine the efficiency of transfer to the membrane and the other half can be used for immunoblotting (i.e., western blotting). Detection limits of each staining method are given along with a list of compatible blot transfer membranes and gels. A support protocol describes a method for alkali treatment that enhances subsequent staining of bound proteins.
在本单元的基本方案和替代方案中,蛋白质从聚丙烯酰胺凝胶电转印至印迹转移膜后进行染色。如果将感兴趣的样品进行一式两份的电泳并转移至印迹转移膜,可将膜的一半进行染色以确定转移至膜上的效率,另一半则用于免疫印迹(即蛋白质印迹法)。每种染色方法的检测限以及兼容的印迹转移膜和凝胶列表一并给出。一个支持方案描述了一种碱处理方法,该方法可增强对结合蛋白质的后续染色效果。