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十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离蛋白质的银染原理与方法

Principle and Method of Silver Staining of Proteins Separated by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis.

作者信息

Kumar Gaurav

机构信息

Oklahoma Medical Research Foundation, University of Oklahoma, Oklahoma City, OK, USA.

出版信息

Methods Mol Biol. 2018;1853:231-236. doi: 10.1007/978-1-4939-8745-0_26.

Abstract

Silver staining is an excellent technique for detecting proteins which are separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis due to its efficiency of detecting proteins present in nanograms. The technique is based on the simple principle that selective reduction of silver into metallic silver occurs at the initiation site in the close proximity of protein molecules. The staining process sequentially consists of protein fixation, sensitization, washing, silver impregnation, and finally development of image. Depending upon the amount of silver incorporated into the protein bands, different color of gel is produced on silver staining. Though different protocols of silver staining exist, the method described here is easy, cheap, reliable, and very sensitive.

摘要

银染是一种用于检测经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离的蛋白质的出色技术,因为它能有效检测出纳克级的蛋白质。该技术基于一个简单的原理,即在蛋白质分子附近的起始位点,银会选择性地还原为金属银。染色过程依次包括蛋白质固定、敏化、洗涤、银浸染,最后显影成像。根据掺入蛋白质条带中的银量,银染后凝胶会呈现出不同的颜色。虽然存在不同的银染方案,但这里描述的方法简单、便宜、可靠且非常灵敏。

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