OTUD5 Antibody / OTU deubiquitinase 5 (FY12167)

Immunofluorescent staining of OTUD5 using anti-OTUD5 antibody (red) and anti-Beta Tubulin antibody (green). OTUD5 was detected in immunocytochemical section of cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-OTUD5 antibody and mouse anti-Beta Tubulin antibody overnight at 4oC. Cy3 Conjugated Goat Anti-Rabbit IgG and FITC Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of OTUD5 using anti-OTUD5 antibody. Lane 1: human 293T whole cell lysates, Lane 2: human Hela whole cell lysates, Lane 3: human Jurkat whole cell lysates, Lane 4: human K562 whole cell lysates, Lane 5: rat liver tissue lysates, Lane 6: rat C6 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-OTUD5 antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. The predicted band size for OTUD5 is at 61 kDa. Phosphorylation causes the protein to migrate more slowly in SDS-PAGE and it is commonly observed at 70-80 kDa.