Protein kinase C (PKC) is a family of serine- and threonine-specific protein kinases that can be activated by calcium and the second messenger diacylglycerol. PKC family members phosphorylate a wide variety of protein targets and are known to be involved in diverse cellular signaling pathways. PKC family members also serve as major receptors for phorbol esters, a class of tumor promoters. Each member of the PKC family has a specific expression profile and is believed to play a distinct role in cells. The protein encoded by this gene is one of the PKC family members. This kinase has been reported to play roles in many different cellular processes, such as cell adhesion, cell transformation, cell cycle checkpoint, and cell volume control. Knockout studies in mice suggest that this kinase may be a fundamental regulator of cardiac contractility and Ca(2+) handling in myocytes.

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Western blotting analysis using Phospho-PKC alpha (T497) antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with Phospho-PKC alpha (T497) antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Western blotting analysis using Phospho-PKC alpha (T497) antibody . 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 Phospho-PKC alpha (T497) antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Flow cytometric analysis of Phospho-PKC alpha (T497) expression in C2C12 cells using Phospho-PKC alpha (T497) antibody . Green, isotype control; red, Phospho-PKC alpha (T497).

Immunocytochemical staining of C2C12 cells with Phospho-PKC alpha (T497) antibody . 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.

Immunocytochemical staining of HT-1080 cells using Phospho-PKC alpha (T497) antibody , 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.

Validation of PKC alpha knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HT1080 cells were stained with Phospho-PKC alpha (T497) antibody and analyzed using BD flow cytometer.

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