HIBADH Antibody / 3-Hydroxyisobutyrate dehydrogenase (FY12781)

Flow Cytometry analysis of cells using anti-HIBADH 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-HIBADH 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 (Red line) was also used as a control.

Western blot analysis of HIBADH using anti-HIBADH antibody. Lane 1: human 293T whole cell lysates, Lane 2: human whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human SIHA whole cell lysates, Lane 5: rat heart tissue lysates, Lane 6: rat NRK whole cell lysates, Lane 7: mouse heart tissue lysates, Lane 8: 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-HIBADH 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. A predominant band at ~32 kDa corresponds to the processed mitochondrial form, with a weaker band near ~35 kDa representing the precursor or partially processed species relative to the ~35 kDa calculated mass.