title: A visual chip-based coimmunoprecipitation technique for analysis of protein-protein interactions.
abstract: 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.
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Chen
authors.forename: Qiongming
authors.initials: Q
authors.lastname: Liu
authors.forename: Zhoumin
authors.initials: Z
authors.lastname: Li
authors.forename: Wenying
authors.initials: W
authors.lastname: Zhong
authors.forename: Wei
authors.initials: W
authors.lastname: He
authors.forename: Danke
authors.initials: D
authors.lastname: Xu
mesh_terms: 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
chemical_list: D000906:Antibodies; D012834:Silver; D006046:Gold; D019809:Streptavidin