Validation of IKK alpha knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with IKK alpha antibody (R020313, 1:2,000) and analyzed using BD flow cytometer.

Immunocytochemical staining of HeLa cells using IKK alpha antibody (R020313, 1:1,000), Top panel: wild-type (WT); Bottom panal: IKK alpha shRNA knockdown (KD). Nuclei were stained blue with DAPI; IKK alpha was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm. Permeabilization: Triton.

Flow cytometric analysis of IKK alpha expression in HepG2 cells using IKK alpha antibody (R020313, 1:2,000). Green, isotype control; red, IKK alpha.

Immunocytochemical staining of HepG2 cells with component of inhibitor of IKK alpha antibody (R020313, 1:1,000). Nuclei were stained blue with DAPI; Component of inhibitor of IKK alpha was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: medium. Scale bar: 20 μm.

Western blotting analysis using IKK alpha antibody (R020313). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with IKK alpha antibody (R020313, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Western blotting analysis using IKK alpha antibody (R020313). IKK alpha expression in wild type (WT) and IKK alpha shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with IKK alpha antibody (R020313, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.