This co-immunoprecipitation assay is a simple and sensitive tool to quickly confirm that two proteins interact when they are expressed in mammalian cells.
The assay requires the following three components:
- The Matchmaker Chemiluminescent Co-IP Vector Set enables you construct and strongly express two tagged fusion proteins.
- The Matchmaker Chemiluminescent Co-IP Kit allows you to co-immunoprecipitate the interacting fusion protein complexes from cell extracts.
- The ProLabel Detection Kit II (Cat. No. 631629) is used to perform a chemiluminescent assay that directly quantifies precipitated protein complexes—no SDS-PAGE or Western blotting is required.
Co-Immunoprecipitation of Two Proteins
Protein 1 cDNAs are cloned in the pAcGFP1-C vector to construct fusion proteins containing the brightly fluorescent Living Colors AcGFP1 tag. This tag enables fusion protein expression to be visualized under fluorescent microscopy and also functions as the antibody epitope used for immunoprecipitation. Protein 2 cDNAs are cloned in the pProLabel-C vector to incorporate the ProLabel tag, which enables sensitive chemiluminescent detection.