Co-Immunoprecipitation Protocol

Co-Immunoprecipitation Protocol Co-Immunoprecipitation (Co-IP) is an extension of immunoprecipitation (IP) with which Co-IP shares the same fundamental principle of the specific antigen-antibody reaction. By targeting a known protein with a specific antibody, it may be possible to pull the entire protein complex out of solution, thereby identifying unknown members of the complex. However, Co-IP requires greater care and more physiologically relevant conditions than traditional IP. When successful, Co-IP pulls down not only the protein of interest but also its interaction partners. Thus, Co-IP is a powerful technique to identify protein complexes and determine protein-protein interactions. Here we provide a detailed procedure for Co-IP to study protein–protein interactions.

Regents: PBS: 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 2 mM KH2PO4, pH to 7.4.

Cell lysis buffer: 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1mM EDTA, 1% NP-40, 1% Na-deoxycholate, 0.1% SDS, sterile-filtered, add protease inhibitor cocktail before use.

3×SDS buffer：150mM Tris-HCl (pH6.8), 6%(W/V) SDS, 0.3%(W/V) BPB, 30% glycerol, 3% β-mercaptoethanol Antibody Equipment: 15-mL Conical tube Refrigerated microcentrifuge Sonicator Magnetic beads Magnetic separation rack https://www.creative-diagnostics.com/co-immunoprecipitation-co-ip-protocol.htm