Anti-Rsk 2/MAPKAP Kinase 1b/RPS6KA3 Picoband Antibody

IF analysis of RPS6KA3 using anti-RPS6KA3 antibody (A02215-2) and anti-Beta Tubulin antibody (M01857-3).
RPS6KA3 was detected in immunocytochemical section of SiHa cell. 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-RPS6KA3 Antibody (A02215-2) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and DyLight®594 Conjugated Goat Anti-Mouse IgG (BA1141) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Immunoprecipitating RPS6KA3 in A549 whole cell lysate .
Western blot analysis of RPS6KA3 using anti-RPS6KA3 antibody (A02215-2).
Lane 1: A549 whole cell lysates (30ug)
Lane 2: Rabbit control IgG instead of anti-RPS6KA3 antibody in A549 whole cell lysate.
Lane 3: anti-RPS6KA3 antibody (2μg) + A549 whole cell lysate (500μg)
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-RPS6KA3 antigen affinity purified polyclonal antibody (A02215-2) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for RPS6KA3 at approximately 84 kDa. The expected band size for RPS6KA3 is at 84 kDa.

Flow Cytometry analysis of 293T cells using anti-RPS6KA3 antibody (A02215-2).
Overlay histogram showing 293T cells stained with A02215-2 (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-RPS6KA3 Antibody (A02215-2, 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.