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蛋白质凝胶的快速考马斯亮蓝染色

Rapid coomassie blue staining of protein gels.

作者信息

Simpson Richard J

出版信息

Cold Spring Harb Protoc. 2010 Apr;2010(4):pdb.prot5413. doi: 10.1101/pdb.prot5413.

Abstract

Coomassie brilliant blue R250 (CBR-250) and silver staining are the most widely used methods for the routine visualization of proteins separated by SDS-PAGE. CBR-250 is an organic dye that complexes with basic amino acids, such as arginine, lysine, and histidine, as well as tyrosine. Conventional CBR-250 staining is capable of detecting as little as 30-100 ng of protein, but sensitivity can be improved by performing the staining and destaining at elevated temperatures. The method described in this protocol, which is a modified version of the conventional Coomassie protocol, speeds up the destaining process for faster results with increased sensitivity and is compatible with mass-spectrometry-based methods for identifying proteins.

摘要

考马斯亮蓝R250(CBR - 250)染色和银染是用于通过SDS - PAGE分离的蛋白质常规可视化的最广泛使用的方法。CBR - 250是一种有机染料,它与碱性氨基酸(如精氨酸、赖氨酸和组氨酸)以及酪氨酸结合。传统的CBR - 250染色能够检测低至30 - 100 ng的蛋白质,但通过在升高的温度下进行染色和脱色可以提高灵敏度。本方案中描述的方法是传统考马斯方法的改进版本,它加快了脱色过程,以更快地获得结果,同时提高了灵敏度,并且与基于质谱的蛋白质鉴定方法兼容。

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