How is the Thermo Scientific Pierce Classic IP Kit method different from traditional immunoprecipitation?
Both traditional and Classic IP methods involve incubation of an antibody with a sample that contains the protein antigen of interest, followed by capture of the antibody:antigen complex to immobilized Protein A or Protein G beaded agarose resin. After washing to remove nonbound (presumably undesired) components of the sample, the antigen and antibody are eluted and separated from the resin. In the traditional method, the components of the IP-complex are eluted by boiling the resin in denaturing sample loading buffer for analysis by SDS-PAGE. The entire procedure is performed in a microcentrifuge tube, requiring that the solution be carefully withdrawn from the agarose resin after they have been pelleted by centrifugation. In the very least, both the target antigen and the antibody are eluted and separated into individual subunits in the final fraction, resulting in the presence of prominent heavy and light chain antibody bands when analyzed by SDS-PAGE. With the Classic IP method, the
