A visual chip-based coimmunoprecipitation technique for analysis of protein-protein interactions.
Here we report a visual chip-based coimmunoprecipitation (vChip-coIP) platform for analysis of protein-protein interactions (PPIs) by combining advantages of an antibody microarray, traditional coIP, and a silver enhancement detection method. The chip was fabricated by spotting anti-Flag antibody on aldehyde-modified slides, and the resulting platform could assay immunoprecipitate from a small amount of crude cell lysates containing Flag-bait and Myc-prey. The interaction signals are visible using biotinylated anti-Myc antibody and colloidal gold-labeled streptavidin followed by a silver enhancement detection method. It is shown that vChip-coIP is a simple, cost-effective, and highly efficient platform for the comprehensive study of PPIs.
Chen
Qing
Q
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China.
Liu
Qiongming
Q
Li
Zhoumin
Z
Zhong
Wenying
W
He
Wei
W
Xu
Danke
D
D000906:Antibodies / Q000737:chemistry* / Q000276:immunology; D019921:Biotinylation; D002460:Cell Line; D006046:Gold / Q000737:chemistry; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D012834:Silver / Q000737:chemistry; D019809:Streptavidin / Q000737:chemistry
D000906:Antibodies; D012834:Silver; D006046:Gold; D019809:Streptavidin