NUDT12 Antibody / Nudix hydrolase 12 (FY12015)

Flow Cytometry analysis of cells using anti-NUDT12 antibody. Overlay histogram showing cells stained with (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NUDT12 antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of NUDT12 using anti-NUDT12 antibody. Lane 1: human 293T whole cell lysates, Lane 2: human Hela whole cell lysates, Lane 3: rat liver tissue lysates, Lane 4: mouse liver tissue lysates, Lane 5: mouse Hepa1-6 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-NUDT12 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. Expected molecular weight of NUDT12 ~52 kDa (462 aa). Observed bands at ~47 kDa (human) and ~49 kDa (mouse) are consistent with published characterizations and vendor validation data that report NUDT12 migrating slightly below 50 kDa. The downward shift may reflect mitochondrial/peroxisomal targeting sequence removal or conformational/gel migration effects.