Flow cytometric analysis of Raf1 expression in HepG2 cells using Raf1 antibody (Cat#61510, 1:2,000). Green, isotype control; red, Raf1.
Immunocytochemical staining of HepG2 cells with Raf1 antibody (Cat#61510, 1:1,000). Nuclei were stained blue with DAPI; Raf1 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-Raf1 antibody (Cat#61510). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with anti-Raf1 antibody (Cat#61510, 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-Raf1 antibody (Cat#61510). Raf1 expression in wild type (WT) and Raf1 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-Raf1 antibody (Cat#61510, 1:5000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Validation of Raf1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with anti-Raf1 antibody (Cat#61510, 1:2,000) and analyzed using CytoFLEX.
Immunocytochemical staining of HeLa cells using anti- Raf1 antibody (Cat#61510, 1:1,000), Top panel: wild-type (WT); Bottom panal: Raf1 shRNA knockdown (KD). Nuclei were stained blue with DAPI; Raf1 was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm. Permeabilization: Triton.
