PTPN11 | SHP-2 (p-Y81) Rabbit

Tissue/cell: Rat brain tissue; 4% P‌‌araformaldehyde-fixed and paraffin-embedded;
_x000D_ 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,C-0005) at 37℃ for 20 min;
_x000D_ Incubation: Anti-Phospho-SHP2(Tyr81)Polyclonal Antibody, Unconjugated(bs-5638R) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
_x000D_

P‌‌araformaldehyde-fixed, paraffin embedded (Mouse brain); 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 (Phospho-SHP2 (Tyr81)) Polyclonal Antibody, Unconjugated (bs-5638R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control（black line）:Hela.
_x000D_ Primary Antibody (green line): Rabbit Anti-Phospho-PTPN11 (Tyr81) antibody (bs-5638R)
_x000D_ Dilution:2ug/Test;
_x000D_ Secondary Antibody（white blue line）: Goat anti-rabbit IgG-FITC
_x000D_ Dilution: 0.5ug/Test.
_x000D_ Isotype control（orange line）: Normal Rabbit IgG
_x000D_ Protocol
_x000D_ The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.