STK17A Antibody / Serine/threonine-protein kinase 17A (FY12534)

Immunoprecipitating STK17A in U251 whole cell lysate. Western blot analysis of STK17A using anti-STK17A antibody. Lane 1: U251 whole cell lysates (30ug), Lane 2: Rabbit control IgG instead of anti-STK17A antibody in U251 whole cell lysate, Lane 3: anti-STK17A antibody (2ug) + U251 whole cell lysate (500ug). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-STK17A antibody at a dilution of 0.5 ug/ml and probed with a mouse anti-rabbit IgG-HRP secondary antibody (Light Chain). The signal is developed using ECL Plus Western Blotting Substrate. STK17A (~47 kDa predicted) was detected as a major band at ~53 kDa, consistent with phosphorylation-dependent migration shift and the acidic C-terminal domain known to cause slower SDS-PAGE mobility.

Western blot analysis of STK17A using anti-STK17A antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human PC-3 whole cell lysates, Lane 2: human U251 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-STK17A 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 an ECL Plus Western Blotting Substrate. STK17A (~47 kDa predicted) was detected as a major band at ~53 kDa, consistent with phosphorylation-dependent migration shift and the acidic C-terminal domain known to cause slower SDS-PAGE mobility.

Flow Cytometry analysis of PC-3 cells using anti-STK17A antibody. Overlay histogram showing PC-3 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-STK17A 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.