NSL1 Antibody / Kinetochore-associated protein NSL1 (FY13016)

Flow Cytometry analysis of HepG2 cells using anti-NSL1 antibody. Overlay histogram showing HepG2 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-NSL1 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 NSL1 using anti-NSL1 antibody. Lane 1: rat thymus tissue lysates, Lane 2: rat RH-35 whole cell lysates, Lane 3: mouse thymus tissue lysates, Lane 4: mouse NIH/3T3 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-NSL1 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 dominant band is detected at ~36 kDa, matching the apparent migration reported for NSL1 on SDS-PAGE (predicted ~32 kDa from its 281-aa sequence). A weaker lower band in mouse (and possibly rat) near the low-30-kDa region may represent limited proteolysis or a minor transcript variant.