All lanes : Anti-PKC theta (phospho T538) antibody [F4H4L1] (ab203565) at 3 µg/mlLane 1 : Jurkat lysates stimulated with 100 ng/mL PMA for 1 hourLane 2 : Jurkat lysates stimulated with 100 ng/mL PMA for 1 hour with Immunogen phosphopeptideLane 3 : Jurkat lysates stimulated with 100 ng/mL PMA for 1 hour with Non-phosphopeptide
Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human breast carcinoma tissue labeling PKC theta (phospho T538) with ab203565 at 5 µg/mL followed by DAB staining. Magnification: 20x.
Immunofluorescence analysis of HeLa cells labeling PKC theta (phospho T538) with ab503565 at 10 μg/mL in the absence of peptides (top panels), presence of phosphopeptide used as immunogen (bottom panels) or non-phosphopeptide (middle panels). Alexa Fluor® 488 goat anti-rabbit used at 1/1000 was used as secondary antibody. Actin was stained with Alexa Fluor® 568 Phalloidin. Hoechst only (left), PKC-θ [pT538] (AF488) signal only (left center), composite image with Phalloidin (right center), and composite image without Hoechst (right).
Flow cytometric analysis of Jurkat cells (incubated with 100 μM PMA for 1 hour prior to being fixed and permeabilized) labeling PKC theta (phospho T538 with ab203565 at 0.1 μg followed by Alexa Fluor® 488 goat anti-rabbit IgG (grey). Pre-incubation with the immunogenic phosphopeptide decreased the signal (red), whereas incubation with the non-phosphopeptide did not (blue).