Fig. 2. Immunoprecipitation of the ryanodine receptor (RyR), protein phosphatase 2A (PP2A) and PKA with mAKAP antiserum. A 13,000 g pellet fraction of whole heart extract was solubilized with 0.5% Triton X-100 and 150 mM NaCl and cleared by repeat centrifugation. The solubilized pellet fraction was used for immunoprecipitation experiments with control preimmune serum (lane 2) or specific mAKAP VO54 antiserum (lane 3). Whole heart extract (5 µg, lane 1) and immunoprecipitates were subjected to SDS-PAGE and immunoblotting with purified mAKAP antibody and specific RyR, PLB, PP2A C-subunit and PKA C-subunit monoclonal antibodies. The migration of the respective protein and molecular weight markers are indicated for each panel. Each panel is representative of three separate experiments.