一种优化的共免疫沉淀方案，用于使用质谱分析细胞系中内源性蛋白质-蛋白质相互作用。
An optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry.
机构信息
Mass Spectrometry and Proteomics Core Facility, University of Nebraska Medical Center, Omaha, NE 68198, USA.
Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
出版信息
STAR Protoc. 2022 Mar 10;3(1):101234. doi: 10.1016/j.xpro.2022.101234. eCollection 2022 Mar 18.
This protocol represents an optimized proteomics-based protocol for the endogenous protein enrichment and protein-protein interaction analysis. This 2-step protocol consists of: 1) co-immunoprecipitation of the bait protein; 2) the bait-protein interactions analysis using LC-MS/MS. Here, we used Dynabeads® for the enrichment of the target protein (the bait) and its interactors. We have tested the protocol using several different cell lines. Our conclusion is that the protocol is applicable to different cell lines and species. For complete details on the use and execution of this protocol, please refer to Lagundžin et al. (2019).
本方案代表了一种优化的基于蛋白质组学的内源性蛋白质富集和蛋白质-蛋白质相互作用分析方案。该 2 步方案包括：1）诱饵蛋白的共免疫沉淀；2）使用 LC-MS/MS 进行诱饵蛋白相互作用分析。在这里，我们使用 Dynabeads®来富集靶蛋白（诱饵）及其相互作用蛋白。我们已经使用几种不同的细胞系测试了该方案。我们的结论是，该方案适用于不同的细胞系和物种。有关该方案使用和执行的完整详细信息，请参考 Lagundžin 等人（2019 年）。
Zotero 插件