Western blotting analysis using protein kinase c alpha antibody (Cat#63197). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with protein kinase c alpha antibody (Cat#63197, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Western blotting analysis using protein kinase c alpha antibody (Cat#63197). Protein kinase c alpha expression in wild type (WT) and protein kinase c alpha (PRKCA) shRNA knockdown (KD) HT-1080 cells with 20 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with protein kinase c alpha antibody (Cat#63197, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Immunohistochemistry was performed on paraffin-embedded human sigmoid colon carcinoma using protein kinase c alpha antibody (Cat#63197, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar, 25 μm.
Immunocytochemical staining of HepG2 cells with protein kinase c alpha antibody(Cat#63197, 1:1000) . Nuclei were stained blue with DAPI; Protein kinase c alpha 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.
Immunocytochemical staining of HT-1080 cells using protein kinase c alpha antibody (Cat#63197, 1:1,000), Top panel: wild-type (WT); Bottom panal: Protein kinase c alpha shRNA knockdown (KD). Nuclei were stained blue with DAPI; Protein kinase c alpha was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.
Flow cytometric analysis of protein kinase c alpha expression in HepG2 cells using protein kinase c alpha antibody (Cat#63197, 1:2,000). Green, isotype control; red, Protein kinase c alpha.
Validation of protein kinase c alpha knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HT-1080 cells were stained with protein kinase c alpha antibody (Cat#63197, 1:2,000) and analyzed using BD flow cytometer.
