ATP1A4 Antibody / ATPase subunit alpha 4 (FY12601)

Flow Cytometry analysis of MCF-7 cells using anti-ATP1A4 antibody. Overlay histogram showing MCF-7 cells stained with (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-ATP1A4 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 ATP1A4 using anti-ATP1A4 antibody. Lane 1: rat C6 whole cell lysates, Lane 2: mouse testis tissue lysates, Lane 3: mouse Raw264.7 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-ATP1A4 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. Western blot probed with anti-ATP1A4 shows a dominant band at ~100 kDa, slightly below the predicted ~114 kDa, consistent with the known anomalous migration of the Na+/K+-ATPase alpha 4 subunit due to its transmembrane structure and partial deglycosylation.