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用咪唑-锌盐对十二烷基硫酸钠聚丙烯酰胺凝胶进行反向染色:未修饰蛋白质的灵敏检测

Reverse staining of sodium dodecyl sulfate polyacrylamide gels by imidazole-zinc salts: sensitive detection of unmodified proteins.

作者信息

Fernandez-Patron C, Castellanos-Serra L, Rodriguez P

机构信息

Center for Genetic Engineering and Biotechnology (CIGB), La Habana, Cuba.

出版信息

Biotechniques. 1992 Apr;12(4):564-73.

PMID:1380251
Abstract

We report here a modification to copper and zinc chloride staining methods. The introduction of a preincubation of the gels, prior to metal staining, with 0.2 M imidazole allows the formation of a homogeneous background for the subsequent precipitation of the metal chelate. The reported imidazole-zinc staining takes minutes, resulting in reproducible staining patterns with only slightly lower sensitivity than silver staining. The method allows efficient recovery of proteins from previously stained gels and is compatible with immunoidentification on Western blots and also with amino acid analysis and NH2-terminal sequence analysis of transferred proteins. A mechanism is proposed to explain the observed improvement in reproducibility and sensitivity of imidazole preincubation to zinc staining.

摘要

我们在此报告对铜和氯化锌染色方法的一种改进。在金属染色之前,先将凝胶与0.2 M咪唑进行预孵育,这能为后续金属螯合物的沉淀形成均匀的背景。所报道的咪唑 - 锌染色只需几分钟,产生的染色模式具有可重复性,其灵敏度仅略低于银染色。该方法能有效地从先前染色的凝胶中回收蛋白质,并且与蛋白质印迹法上的免疫鉴定兼容,也与转移蛋白质的氨基酸分析和氨基末端序列分析兼容。我们提出了一种机制来解释观察到的咪唑预孵育对锌染色的重复性和灵敏度的改善。

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