Western blotting analysis using anti-phosphoglycerate dehydrogenase antibody (Cat#65056). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with anti-phosphoglycerate dehydrogenase antibody (Cat#65056, 1:2,500) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using anti-phosphoglycerate dehydrogenase antibody (Cat#65056). Phosphoglycerate dehydrogenase expression in wild-type (WT) and phosphoglycerate dehydrogenase (PHGDH) shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with anti-phosphoglycerate dehydrogenase antibody (Cat#65056, 1:2,500) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Validation of Phosphoglycerate dehydrogenase knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with anti-Phosphoglycerate dehydrogenase antibody (Cat#65056, 1:2,000) and analyzed using BD flow cytometer.
Immunocytochemical staining of HeLa cells using anti-Phosphoglycerate dehydrogenase antibody (Cat#65056, 1:1,000), Top panel: wild-type (WT); Bottom panal: Phosphoglycerate dehydrogenase shRNA knockdown (KD). Nuclei were stained blue with DAPI; Phosphoglycerate dehydrogenase was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.
