An optimized co-immunoprecipitation protocol for the analysis of endogenous protein-protein interactions in cell lines using mass spectrometry

被引：6

作者：

Lagundzin, Dragana ^{[1
]}

Krieger, Kimiko L. ^{[2
]}

Law, Henry C. -H. ^{[3
]}

Woods, Nicholas T. ^{[3
]}

机构：

[1] Univ Nebraska Med Ctr, Mass Spectrometry & Prote Core Facil, Omaha, NE 68198 USA

[2] Baylor Coll Med, Dept Mol & Cellular Biol, Houston, TX 77030 USA

[3] Univ Nebraska Med Ctr, Eppley Inst Res Canc & Allied Dis, Fred & Pamela Buffett Canc Ctr, Omaha, NE 68198 USA

来源：

STAR PROTOCOLS | 2022年 / 3卷 / 01期

关键词：

Mass Spectrometry; Protein Biochemistry; Protein expression and purification; Proteomics;

D O I：

10.1016/j.xpro.2022.101234

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 DynabeadsO 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 Lagundz & DBLBOND;in et al. (2019).

引用

页数：14

共 7 条

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