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一种将考马斯亮蓝染色的蛋白质从聚丙烯酰胺凝胶转移至透明胶片上的便捷方法。

A convenient procedure for transfer blotting of coomassie blue stained proteins from PAGE gels to transparencies.

作者信息

Chu Y H, Whitesides G M

机构信息

Department of Chemistry, Harvard University, Cambridge, MA 02138.

出版信息

Biotechniques. 1993 Jun;14(6):925-30.

PMID:7687447
Abstract

Proteins stained with Coomassie Brilliant Blue I were transferred effectively by blotting from polyacrylamide gel electrophoresis (PAGE) gels to transparencies of the type used in plain-paper copiers. The details of the original electropherogram were retained on transfer and did not fade over a period of three years. Both the protein and the associated dye transfer; however, protein does not transfer in the absence of dye. Protein patterns present in several types of gels--sodium dodecyl sulfate (SDS)-PAGE, non-denaturing PAGE, isoelectric focusing PAGE and SDS-agarose--all transferred successfully after staining with dye 1. Proteins visualized with other organic dyes such as Fast Green FCF 2, Uniblue A 3 and Procion Blue MX-R 4 also transferred, but proteins stained with Stains-all 5 or silver did not. This transfer provides a simple, economical way to preserve data from slab gel electrophoresis and a convenient method to display data using an overhead projector. The blotted transparencies are also excellent substrates for use with gel scanning densitometers.

摘要

用考马斯亮蓝I染色的蛋白质通过印迹法能有效地从聚丙烯酰胺凝胶电泳(PAGE)凝胶转移至普通纸复印机所用类型的透明胶片上。原始电泳图谱的细节在转移后得以保留,并且在三年时间内都没有褪色。蛋白质和相关染料都能转移;然而,在没有染料的情况下蛋白质不会转移。存在于几种类型凝胶中的蛋白质模式——十二烷基硫酸钠(SDS)-PAGE、非变性PAGE、等电聚焦PAGE和SDS-琼脂糖——在用染料1染色后都成功转移。用其他有机染料如固绿FCF 2、优尼蓝A 3和普施安蓝MX-R 4可视化的蛋白质也能转移,但用全染剂5或银染色的蛋白质不能转移。这种转移提供了一种简单、经济的方法来保存平板凝胶电泳的数据,以及一种使用投影仪显示数据的便捷方法。印迹后的透明胶片也是与凝胶扫描密度计配合使用的优质底物。

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