TAOK1 Antibody / TAO kinase 1 (FY12950)

Immunoprecipitating TAOK1 in Hela whole cell lysate. Western blot analysis of TAOK1 using anti-TAOK1 antibody. Lane 1: Hela whole cell lysates (30ug), Lane 2: Rabbit control IgG instead of anti-TAOK1 antibody in Hela whole cell lysate, Lane 3: anti-TAOK1 antibody (2ug) + Hela whole cell lysate (500ug). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-TAOK1 antibody at a dilution of 0.5 ug/ml and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. A predominant band is detected at ~130 kDa, higher than the ~116 kDa prediction. The upward shift is consistent with hyperphosphorylated TAOK1, which migrates slower on SDS-PAGE.

Western blot analysis of TAOK1 using anti-TAOK1 antibody. Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human Hela whole cell lysates, Lane 2: human whole cell lysates, Lane 3: human RT4 whole cell lysates, Lane 4: human SIHA whole cell lysates, Lane 5: rat testis tissue lysates, Lane 6: rat brain tissue lysates, Lane 7: mouse testis tissue lysates, Lane 8: mouse brain tissue 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-TAOK1 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. A predominant band is detected at ~130 kDa, higher than the ~116 kDa prediction. The upward shift is consistent with hyperphosphorylated TAOK1, which migrates slower on SDS-PAGE.

Flow Cytometry analysis of HeLa cells using anti-TAOK1 antibody. Overlay histogram showing HeLa 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-TAOK1 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.