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relaxin 10 months ago

If you use antibody against a bait protein to perform the first co-immunoprecipitation and identify its binding partner on a Western blot using antibody against the binding partner, then the reverse co-immunoprecipitation will be to perform the immunoprecipitation with an antibody against the novel binding partner, and probe the immunoblot with an antibody against the original bait protein of interest. This is a confirmation step to rule out nonspecific binding between these two proteins with one of antibodies.