Western blot analysis was performed on whole cell extracts (30 µg lysate) of HCT 116 (Lane 1), Caco-2 (Lane 2), COLO 205 (Lane 3), HT-29 (Lane 4), U-2 OS (Lane 5), HEK-293 (Lane 6) ,Hep G2 (Lane 7) and MCF-7 (Lane 8). The blots were probed with Anti-ING2 Recombinant Rabbit Polyclonal Antibody (Product # 711779, 1:200 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 32 kDa band corresponding to ING2 was observed across the cell lines tested.
Knockdown of ING2 was achieved by transfecting MCF7 cells with ING2 specific siRNA (Silencer® select Product # s7433 and Product # s7432). Western blot analysis (Fig a) was performed using whole cell extracts from the MCF-7 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blots were probed with Anti-ING2 Recombinant Rabbit Polyclonal Antibody (Product # 711779, 1:200 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 histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to ING2.
Supplier Page from Thermo Fisher Scientific for ING2 Antibody