Western blotting analysis using OTUB1 antibody (Cat#63556). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with OTUB1 antibody (Cat#63556, 1:5,000) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using OTUB1 antibody (Cat#63556). OTUB1 expression in wild type (WT) and OTUB1 shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with OTUB1 antibody (Cat#63556, 1:5,000) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Immunocytochemical staining of HAP-1 cells with OTUB1 antibody (Cat#63556, 1:1,000). Nuclei were stained blue with DAPI; OTUB1 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Scale bar, 20 μm.
Flow cytometric analysis of OTUB1 expression in HAP-1 cells using OTUB1 antibody (Cat#63556, 1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 100% Methanol, chill on -20℃ before use. Green, isotype control; red, OTUB1.
Validation of OTUB1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with OTUB1 antibody (Cat#63556, 1:2,000) and analyzed using BD flow cytometer. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 100% Methanol, chill on -20℃ before use.
