Flow cytometric analysis of 14-3-3 gamma expression in HepG2 cells using 14-3-3 gamma antibody (Cat#61127, 1:2,000). Green, isotype control; red, 14-3-3 gamma.
Immunocytochemical staining of HepG2 cells with 14-3-3 gamma antibody (Cat#61127, 1:1,000). Nuclei were stained blue with DAPI; 14-3-3 gamma 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-14-3-3 gamma antibody (Cat#61127). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with anti-14-3-3 gamma antibody (Cat#61127, 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-14-3-3 gamma antibody (Cat#61127). 14-3-3 gamma expression in wild type (WT) and 14-3-3 gamma shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with anti-14-3-3 gamma antibody (Cat#61127, 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 14-3-3 gamma knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with anti-14-3-3 gamma antibody (Cat#61127, 1:2,000) and analyzed using BD flow cytometer.
