Flow cytometric analysis of Phospho-PKC alpha (T497) expression in C2C12 cells using Phospho-PKC alpha (T497) antibody (Cat#61850, 1:2,000). Green, isotype control; red, Phospho-PKC alpha (T497).
Immunocytochemical staining of C2C12 cells with anti-Phospho-PKC alpha (T497) antibody (Cat#61850, 1:1,000) . Nuclei were stained blue with DAPI; Phospho-PKC alpha (T497) was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Scale bar, 20 μm.
Western blotting analysis using anti-Phospho-PKC alpha (T497) antibody (Cat#61850). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with anti-Phospho-PKC alpha (T497) antibody (Cat#61850, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using anti-Phospho-PKC alpha (T497) antibody (Cat#61850). Phospho-PKC alpha (T497) expression in wild type (WT) and Phospho-PKC alpha (T497) shRNA knockdown (KD) HT-1080 cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with anti-Phospho-PKC alpha (T497) antibody (Cat#61850, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Validation of PKC alpha knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HT1080 cells were stained with anti-Phospho-PKC alpha (T497) antibody (Cat#61850, 1:2,000) and analyzed using BD flow cytometer.
Immunocytochemical staining of HT-1080 cells using anti-Phospho-PKC alpha (T497) antibody (Cat#61850, 1:1,000), Top panel: wild-type (WT); Bottom panal: Phospho-PKC alpha (T497) shRNA knockdown (KD). Nuclei were stained blue with DAPI; Phospho-PKC alpha (T497) was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.
