Protocol to test for the formation of ternary protein complexes in vivo or in vitro using a two-step
Bo-Wen Zhang1, Zi-Juan Wei1, Qian-Qian Lou1
1Laboratory of Molecular and Cellular Biology, Department of Cell Biology and Genetics of School of Basic Medical Sciences and Department of General Surgery of Huaihe Hospital, Henan University, Kaifeng, Henan Province 475004, P.R. China; Zhongzhou Laboratory, Zhengzhou, Henan Province 450046, P.R. China.
Abstract:
Co-immunoprecipitation (coIP) is an experimental technique to study protein-protein interactions (PPIs). However, single-step coIP can only be used to identify the interaction between two proteins and does not solve the interaction testing of ternary complexes. Here, we present a protocol to test for the formation of ternary protein complexes in vivo or in vitro using a two-step coIP approach. We describe steps for cell culture and transfection, elution of target proteins, and two-step coIP including western blot analyses. For complete details on the use and execution of this protocol, please refer to Li et al.1.
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