Western blotting analysis using ALKBH1 antibody (Cat#69441). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with ALKBH1 antibody (Cat#69441, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using ALKBH1 antibody (Cat#69441). ALKBH1 expression in wild type (WT) and ALKBH1 shRNA knockdown (KD) Hela cells with 30 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with ALKBH1 antibody (Cat#69441, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Flow cytometric analysis of ALKBH1 expression in HepG2 cells using ALKBH1 antibody (Cat #69441, 1:2,000). Green, isotype control; red, ALKBH1.
Validation of ALKBH1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with ALKBH1 antibody (Cat#69441, 1:2,000) and analyzed using BD flow cytometer.
Immunocytochemical staining of HepG2 cells with ALKBH1 antibody (Cat#69441, 1:1,000) . Nuclei were stained blue with DAPI; ALKBH1 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.
Immunohistochemistry was performed on paraffin-embedded mouse kidney using ALKBH1 antibody (Cat#69441, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar, 25 μm.
