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另一种用于检测聚丙烯酰胺凝胶中蛋白质的改良银染方法。

Yet another improved silver staining method for the detection of proteins in polyacrylamide gels.

作者信息

Tunón P, Johansson K E

出版信息

J Biochem Biophys Methods. 1984 May;9(2):171-9. doi: 10.1016/0165-022x(84)90008-3.

Abstract

Silver staining is very sensitive for detection of proteins in polyacrylamide gels and different procedures have been published. By combining and modifying some of the recipes, a very reproducible method, which is based upon staining with diamine complexes of silver, has been developed. The background staining is negligible and reduced silver does not precipitate on the gel surface. The technique works very well for sodium dodecyl sulfate-polyacrylamide gel electrophoresis in both homogeneous and in gradient gels as well as for two-dimensional (2-D) PAGE. It was possible to detect 1-10 ng of protein corresponding to approximately 50 pg/mm2, provided that a discontinuous buffer system was used, which gives sharp bands.

摘要

银染对检测聚丙烯酰胺凝胶中的蛋白质非常灵敏,已有多种不同的方法发表。通过结合并修改一些配方,开发出了一种基于银二胺络合物染色的、具有高度可重复性的方法。背景染色可忽略不计,还原银不会在凝胶表面沉淀。该技术在均匀凝胶和梯度凝胶的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳以及二维(2-D)聚丙烯酰胺凝胶电泳中均表现良好。如果使用能产生清晰条带的不连续缓冲系统,能够检测到1-10 ng的蛋白质,相当于约50 pg/mm²。

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