FIGURE 2.
Immunoisolation of CPT1a and proteins associated with CPT1a. A, upper panel, SDS-PAGE separation and silver staining of proteins immunoprecipitated (IP) with affinity-purified anti-CPT1a antibodies coupled to protein G beads; lower panel, immunoblot analysis of the starting material (initial MOM solubilized with SDS sample buffer (i-SDS) and initial MOM solubilized with 2% Lubrol PX (i-L-PX)), the pellet after Lubrol PX extraction (Airfuge; P), and the Lubrol PX supernatant (Airfuge; SN) with affinity-purified anti-CPT1a antibodies. Arrows and arrowheads show CPT1a and anti-CPT1a antibodies, respectively. B, immunocapture (IC) of CPT1a and associated proteins with a mixture of five affinity-purified anti-CPT1a antibodies directly coupled to CarboLink. In the eluate, CPT1a, but not anti-CPT1a antibodies, was detected. FT, flow-through with either protein G or CarboLink; E1 and E2, first and second eluents from either protein G- or CarboLink-coupled IgG, respectively.