Flow cytometric analysis of MADD expression in HeLa cells using MADD antibody (R021087, 1:2,000). Green, isotype control; red, MADD.

Immunocytochemical staining of HeLa cells with MADD antibody (R021087, 1:1,000). Nuclei were stained blue with DAPI; MADD 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 MADD antibody (R021087). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with MADD antibody (R021087, 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 MADD antibody (R021087). MADD expression in wild type (WT) and MADD shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with MADD antibody (R021087, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Validation of MADD knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with MADD antibody (R021087, 1:2,000) and analyzed using BD flow cytometer.