Tissue/cell: mouse stomach tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-Phospho-Met (Tyr1234+Tyr1235) Polyclonal Antibody, Unconjugated(SL3272R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Blank control (blue line): U251(fixed with 70% ethanol overnight at 4℃. Cells stained with Primary Antibody for 30 min at room temperature).
Primary Antibody (green line): Rabbit Anti- Phospho-Met (Tyr1234 + Tyr1235) antibody (SL3272R),Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE,Dilution: 1μg /test.