Characterization of the polyclonal anti–γ-adaptin antibody. (A) Postnuclear supernatants (lane 1) or total membranes  (lane 2) were prepared from mouse fibroblasts. The samples were  fractionated by SDS-PAGE, transferred to nitrocellulose, and  analyzed by Western blotting using a polyclonal antibody against  a peptide corresponding to the trunk region (1573; right) or  against the hinge region of the mouse γ-adaptin (γ-H; left). (Arrow) γ-Adaptin; (arrowhead) a 120-kD contaminant. (B) γ-Adaptin  was immunoprecipitated from denaturated (left) or native (right)  HeLa cell lysates with either the polyclonal antibody (1573) or  with the mAb 100/3 as described in Materials and Methods. Both  the immunoprecipitated material (P) and a fraction (10%) of the  supernatant of the immunoprecipitations (S) were analyzed by  Western blotting with either the 1573 or the 100/3 antibodies.