用于分析蛋白质-蛋白质相互作用的共免疫沉淀和半定量免疫印迹。

Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, TX 75390, USA.

Department of Cell Biology, UT Southwestern Medical Center, Dallas, TX 75390, USA.

STAR Protoc. 2021 Jul 6;2(3):100644. doi: 10.1016/j.xpro.2021.100644. eCollection 2021 Sep 17.

Co-immunoprecipitation (co-IP) of protein complexes from cell lysates is widely used to study protein-protein interactions. However, establishing robust co-IP assays often involves considerable optimization. Moreover, co-IP results are frequently presented in non-quantitative ways. This protocol presents an optimized co-IP workflow with an analysis based on semi-quantitative immunoblot densitometry to increase reliability and reproducibility. For complete details on the use and execution of this protocol, please refer to Burckhardt et al. (2021).

从细胞裂解物中免疫共沉淀（co-IP）蛋白质复合物广泛用于研究蛋白质-蛋白质相互作用。然而，建立稳健的 co-IP 检测通常需要相当多的优化。此外，co-IP 结果通常以非定量的方式呈现。本方案介绍了一种优化的 co-IP 工作流程，该流程基于半定量免疫印迹密度测定法进行分析，以提高可靠性和可重复性。有关该方案使用和执行的完整详细信息，请参阅 Burckhardt 等人（2021 年）。