Protein Membrane Overlay Assay: A Protocol to Test Interaction Between Soluble and Insoluble Proteins <em>in vitro</em>

Shoko Ueki; Beno&#238;t Lacroix; Vitaly Citovsky

doi:10.3791/2961

JoVE Journal
Biology

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Protein Membrane Overlay Assay: A Protocol to Test Interaction Between Soluble and Insoluble Proteins in vitro

蛋白膜覆盖含量:一个协议测试可溶性和不溶性蛋白质之间的相互作用在体外

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08:38 min

August 14, 2011

DOI: 10.3791/2961-v

Shoko Ueki¹, Benoît Lacroix¹, Vitaly Citovsky¹

¹Department of Biochemistry and Cell Biology,State University of New York

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测试是必不可少的蛋白质 - 蛋白质相互作用蛋白质功能的夹层。在这里，我们引入

检测蛋白质。蛋白质相互作用对于蛋白质功能的剖析是必不可少的。本视频介绍了一种体外蛋白结合测定法，用于测试膜和固定化蛋白与可溶性蛋白之间的相互作用。

首先，克隆、表达和提取融合蛋白。标记的不溶性蛋白固定在硝酸纤维素膜上，并经过处理以重新折叠至其天然确认。然后用标记的可溶性蛋白探测膜，并进行免疫印迹以检测蛋白质。

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分子生物学 54期蛋白质相互作用叠加在体外培养免疫印迹硝酸纤维素膜不溶性蛋白质