免疫沉淀
蛋白质-蛋白质相互作用
计算生物学
化学
细胞生物学
功能(生物学)
靶蛋白
双杂交筛选
蛋白质功能
生物
酵母
生物化学
基因
作者
Jer-Sheng Lin,Erh‐Min Lai
出处
期刊:Methods in molecular biology
日期:2017-01-01
卷期号:: 211-219
被引量:138
标识
DOI:10.1007/978-1-4939-7033-9_17
摘要
Proteins often do not function as single substances but rather as team players in a dynamic network. Growing evidence shows that protein–protein interactions are crucial in many biological processes in living cells. Genetic (such as yeast two-hybrid, Y2H) and biochemical (such as co-immunoprecipitation, co-IP) methods are the methods commonly used at the beginning of a study to identify the interacting proteins. Immunoprecipitation (IP), a method using a target protein-specific antibody in conjunction with Protein A/G affinity beads, is a powerful tool to identify molecules that interact with specific proteins. Therefore, co-IP is considered to be one of the standard methods of identifying or confirming the occurrence of protein–protein interaction events in vivo. Co-IP experiments can identify proteins via direct or indirect interactions or in a protein complex. Here, we use Agrobacterium type VI secretion system (T6SS) sheath components TssB-TssC41 interaction as an example to describe the principle, procedure, and experimental problems of co-IP.
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