High Performance Immunoprecipitation (HPIP), Indirect method

In biomedical research, it is often necessary to isolate specific proteins from natural sources in order to study them. One of the most commonly used methods to isolate proteins from their biological sources is immunoprecipitation (IP). In IP, proteins from cellular lysate, serum, or other biological fluids are bound to a protein A (ProA) or protein G (ProG) affinity resin using a bridging antibody. The antibody’s antigen binding site binds to the protein of interest, while the Fc chain of the antibody binds to ProA or ProG. After washing nonspecifically bound proteins away, the antibody-protein complex is eluted from the resin prior to further analysis.