Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum tissue); 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 (c-Kit (Tyr730)) Polyclonal Antibody, Unconjugated (SL12914R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Blank control: Hela(blue), the cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with ice-cold 90% methanol for 30 min on ice.
Isotype Control Antibody: Rabbit IgG(orange) ;
Secondary Antibody: Goat anti-rabbit IgG-PE(white blue),
Dilution: 1:200 in 1 X PBS containing 0.5% BSA ;
Primary Antibody Dilution: 1μg in 100 μL1X PBS containing 0.5% BSA(green).