Paraformaldehyde-fixed, paraffin embedded (Rat colon); 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 (Beta catenin) Polyclonal Antibody, Unconjugated (SL20599R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Blank control(blue): Hela (fixed with 2% paraformaldehyde (10 min) , then permeabilized with 0.3%tritionx-100 for 5 min at room temperature).
Primary Antibody:Rabbit Anti-Beta catenin antibody (SL20599R); Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions );
Secondary Antibody: Goat anti-rabbit IgG-FITC), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.