Co-immunoprecipitation from Transfected Cells
Co-immunoprecipitation from Transfected Cells
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DOI：
10.1007/978-1-4939-2425-7_25
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发表时间：
2015-01-01
期刊：
影响因子：
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通讯作者：
Takahashi, Yoshinori
中科院分区：
文献类型：
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作者：
Takahashi, Yoshinori
共免疫沉淀（Co-IP）是最广泛使用的方法之一，用于鉴定与感兴趣的蛋白质相关的新蛋白质或确定已知蛋白质之间的复合物形成。对于这种技术，使用特定的抗体捕获感兴趣的蛋白质。然后使用树脂（免疫沉淀，IP）沉淀抗体结合蛋白，以及与目标蛋白结合的任何蛋白。然后通过一系列洗涤从样品中去除未与目标蛋白质结合的蛋白质。然后用免疫印迹法分析所得的免疫复合物。由于蛋白质-蛋白质相互作用的要求不同，检验不同感兴趣的蛋白质的相互作用伙伴的最佳实验条件必须确定经验。一旦适当的实验条件已经建立，IP/Co-IP程序是简单和直接的。在本章中，讨论了IP/co-IP的标准协议，以及IP/co-IP分析成功的几个关键因素。
Co-immunoprecipitation (Co-IP) is one of the most widely used methods to identify novel proteins that associate with a protein of interest or to determine complex formation between known proteins. For this technique, a protein of interest is captured using a specific antibody. The antibody-bound protein, as well as any proteins bound to the protein of interest, is then precipitated using a resin (immunoprecipitation, IP). Proteins that are not bound to the protein of interest are then removed from the sample with a series of washes. The resulting immunocomplexes are then analyzed by immunoblot. As the requirements for protein-protein interactions vary, optimal experimental conditions for examining the interacting partners of different proteins of interest must be determined empirically. Once appropriate experimental conditions have been established, the IP/Co-IP procedure is simple and straightforward. In this chapter, a standard protocol for IP/co-IP, with several key factors for the success of IP/co-IP analyses, is discussed.