Knockdown of E6AP was achieved by transfecting HeLa cells with E6AP specific validated siRNA (Silencer® select Product # s14605). Western blot analysis (Fig a) was performed using modified whole cell extracts (1% SDS) from E6AP knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Anti-E6AP Recombinant Rabbit Monoclonal Antibody (Product # 703785, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in the histogram (Fig b). Loss of signal upon siRNA mediated knockdown confirms that antibody is specific to E6AP.