SIRT3 encodes a member of the sirtuin family of class III histone deacetylases, homologs to the yeast Sir2 protein. The encoded protein is found exclusively in mitochondria, where it can eliminate reactive oxygen species, inhibit apoptosis, and prevent the formation of cancer cells. SIRT3 has far-reaching effects on nuclear gene expression, cancer, cardiovascular disease, neuroprotection, aging, and metabolic control.

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Western blotting analysis using sirtuin 3 antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with sirtuin 3 antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using NaQ™ ECL Substrate Kit .

Western blotting analysis using sirtuin 3 antibody . Sirtuin 3 expression in wild-type (WT) and sirtuin 3 (SIRT3) shRNA knockdown (KD) HeLa cells with 25 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with sirtuin 3 antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Immunocytochemical staining of HAP-1 cells with Sirtuin 3 antibody . Nuclei were stained blue with DAPI; Sirtuin 3 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.

Validation of Sirtuin 3 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Sirtuin 3 antibody and analyzed using BD flow cytometer.

Immunocytochemical staining of HeLa cells using Sirtuin 3 antibody , Top panel: wild-type (WT); Bottom panal: Sirtuin 3 shRNA knockdown (KD). Nuclei were stained blue with DAPI; Acyl-CoA dehydrogenase short chain was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.

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