A procedure for the detection of free thiol-containing proteins on a polyvinylidene difluoride membrane.

T Kamiya
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引用次数: 1

Abstract

Bovine serum albumin (BSA), which has a free thiol, was blotted onto a polyvinylidene difluoride membrane. The membrane was reacted with a sulfhydryl-reactive (maleimide-containing) biotin derivative, 1-biotinamido-4-[4'-(maleimidomethyl) cyclohexanecarboxamido] butane (Biotin-BMCC), and then probed. BSA on membranes was detected semi-quantitatively at 50 ng of protein (0.76 pmol of free thiol) and among range of higher extent. BSA on membranes was less efficiently biotinylated compared with biotinylation in solution. Regardless, these results suggested that sulfhydryl-containing proteins were specifically and semi-quantitatively identified by membrane biotinylation with Biotin-BMCC.

用于检测聚偏二氟乙烯膜上的游离含硫醇蛋白质的程序。
牛血清白蛋白(BSA),其中有一个自由的硫醇,被印迹到聚偏二氟乙烯膜。该膜与巯基反应性(含马来酰亚胺)生物素衍生物1-生物胺酰胺-4-[4'-(马来酰亚胺甲基)环己烷羧基氨基]丁烷(biotin - bmcc)反应,然后进行探针探测。50 ng蛋白(0.76 pmol游离硫醇)半定量检测膜上的牛血清白蛋白。与溶液中的生物素化相比,膜上的牛白蛋白生物素化效率较低。无论如何,这些结果表明含有巯基的蛋白质可以通过生物素- bmcc的膜生物素化来特异性和半定量地鉴定。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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