Anti-PDK2 Antibody Picoband®

Western blot analysis of PDK2 using anti-PDK2 antibody (PA1994).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat heart tissue lysates,
Lane 2: rat skeletal muscle tissue lysates,
Lane 3: rat brain tissue lysates,
Lane 4: rat kidney tissue lysates,
Lane 5: mouse heart tissue lysates,
Lane 6: mouse skeletal muscle tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse kidney 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-PDK2 antigen affinity purified polyclonal antibody (PA1994) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for PDK2 at approximately 46 kDa. The expected band size for PDK2 is at 46 kDa.

IF analysis of PDK2 using anti-PDK2 antibody (PA1994).
PDK2 was detected in an immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-PDK2 Antibody (PA1994) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Immunoprecipitating (IP) PDK2 in Jurkat whole cell lysate.
Western blot analysis of PDK2 using anti-PDK2 antibody (PA1994);
Lane 1: Jurkat whole cell lysates (30ug);
Lane 2: Rabbit control IgG instead of anti-PDK2 antibody in Jurkat whole cell lysate;
Lane 3: anti-PDK2 antibody (2μg) + Jurkat whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-PDK2 antigen affinity purified polyclonal antibody (PA1994) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BM2007). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for PDK2 at approximately 46 kDa. The expected band size for PDK2 is at 46 kDa.

Flow Cytometry analysis of PC-3 cells using anti-PDK2 antibody (PA1994).
Overlay histogram showing PC-3 cells stained with PA1994 (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-PDK2 Antibody (PA1994, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.