Western blotting analysis using EGFR antibody (Cat#69236). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with EGFR antibody (Cat#69236, 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 EGFR antibody (Cat#69236). EGFR expression in wild type (WT) and EGFR shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with EGFR antibody (Cat#69236, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:50,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Immunocytochemical staining of HepG2 cells with EGFR antibody (Cat#69236, 1:1,000). Nuclei were stained blue with DAPI; EGFR was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: High. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Scale bar: 20 μm.
Flow cytometric analysis of EGFR expression in HepG2 cells using EGFR antibody (Cat#69236,1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 100% Methanol, chill on -20℃ before use. Green, isotype control; red, EGFR.
Validation of EGFR knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HepG2 cells were stained with EGFR antibody (Cat#69236, 1:2,000) and analyzed using BD flow cytometer. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 100% Methanol, chill on -20℃ before use.
