Western blot analysis was performed on whole cell extracts (30 µg lysate) of MDA-MB-231 (Lane 1), SK-BR-3 (Lane 2), T-47D (Lane 3), HeLa (Lane 4), RAW 264.7 (Lane 5), HEK-293 (Lane 6), A-431 (Lane 7) and NIH3T3 (Lane 8). The blot was probed with Anti-TAB1 Recombinant Rabbit Monoclonal Antibody (Product # 703255, 1:500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 1:5000 dilution). A ~55 kDa band corresponding to TAB1 was observed across the cell lines tested.
Knockdown of TAB1 was achieved by transfecting HeLa cells with TAB1 specific validated siRNA (Silencer® select Product # s20451). Western blot analysis (Fig a) was performed using whole cell extracts from the TAB1 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Anti-TAB1 Recombinant Rabbit Monoclonal Antibody (Product # 703255, 1:500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 1:5000 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 TAB1.
Supplier Page from Thermo Fisher Scientific for TAB1 Antibody