EGFR Rabbit Polyclonal Antibody

Blank control (black line): A431. Primary Antibody (green line): Rabbit Anti-EGFR antibody (orb156708), Dilution: 1 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488, Dilution: 0.5 ug/Test. Isotype control (orange line): Normal Rabbit IgG, Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (Human brain glioma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (EGFR) Polyclonal Antibody, Unconjugated (orb156708) at 1:400 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868589) for 90 minutes, and DAPI for nuclei staining.

Sample: NIH/3T3 (Mouse) Cell Lysate at 40 ug, Primary: Anti-EGFR (orb156708) at 1/500 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 170 kD, Observed band size: 181/130 kD.