Department of Pathology and Immunology, Washington University in St. Louis School of Medicine, St. Louis, MO, USA.
Methods Mol Biol. 2022;2444:29-41. doi: 10.1007/978-1-0716-2063-2_3.
Immunoaffinity purification allows for the purification of epitope-tagged proteins and their associated multisubunit complexes from mammalian cells. Subsequent identification of the proteins by proteomic analysis enables unbiased biochemical characterization of their associated partners, potentially revealing the physiological or functional context of any given protein. Here, we use immunoaffinity isolation of the Activating Signal Co-integrator Complex (ASCC) from human cells as an example, demonstrating the utility of the approach in revealing protein complexes involved in genotoxic stress responses.
免疫亲和纯化可从哺乳动物细胞中纯化表位标记蛋白及其相关多亚基复合物。随后通过蛋白质组学分析鉴定这些蛋白质,可以对其相关伴侣进行无偏生化特性分析,从而揭示任何给定蛋白质的生理或功能背景。本文以人细胞中激活信号共整合体复合物(ASCC)的免疫亲和分离为例,展示了该方法在揭示参与遗传毒性应激反应的蛋白质复合物方面的应用。