VALIDATION IMAGES
Formalin-fixed and paraffin-embedded human lung carcinoma labeled with EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Paraformaldehyde-fixed, paraffin embedded human brain glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.
Tissue/cell: Human Rectal Carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15min; Blocking buffer (normal goat serum) at 37℃ for 20 min; Incubation: EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated (bs-0295G-Cy3) used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml, blue) was used to stain the cell nuclei.
Human HUVEC Cells were fixed with 2% PFA (10min at room temperature) and then permeabilized with 90% ice-cold methanol for 30min on ice. The cells were then incubated in 0.5% BSA + 10% goat serum to block non-specific protein-protein interactions for 15 min. Cells were stained with EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:100 for 30 min on ice. The secondary antibody, Goat anti-rabbit IgG-PE, was used for 30 min on ice. Primary Antibody staining (green) is compared to unstained cells (dark blue), secondary only (light blue) and isotype control (orange).
Human A431 Cells were fixed with 2% PFA (10min at room temperature) and then permeabilized with 90% ice-cold methanol for 30min on ice. The cells were then incubated in 2% BSA + 10% goat serum to block non-specific protein-protein interactions for 15 min at room temperature. Cells were stained with EGFR Polyclonal Antibody, Unconjugated (bs-0165R) at 1:33 for 30 min at room temperature. The secondary antibody, Goat anti-rabbit IgG-FITC, was used for 40 min at room temperature. Primary Antibody staining (green) is compared to unstained cells (dark blue), secondary only (light blue) and isotype control (orange).
Paraformaldehyde-fixed, paraffin embedded (rat placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (EGFR) Polyclonal Antibody, Unconjugated (bs-0165R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (EGFR) Polyclonal Antibody, Unconjugated (bs-0165R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.