Estrogen Receptor alpha Rabbit Polyclonal Antibody

Blank control (blue line): MCF7 (blue). Primary Antibody (green line): Rabbit Anti-Estrogen Receptor alpha antibody (orb10602), Dilution: 0.2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 µg/Test. Protocol, The cells were fixed with 80% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (rat uterus), 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 (Estrogen Receptor alpha) Polyclonal Antibody, Unconjugated (orb10602) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Brain (Rat) lysate at 30 ug, Primary: Anti-ER-alpha/beta (orb10602) at 1:200 dilution, Secondary: HRP conjugated Goat Anti-Rabbit IgG (orb572747) at 1:3000 dilution (DAB staining), Predicted band size: 67kD, Observed band size: 67kD.