Rabbit anti-Phospho-β Catenin(Tyr333) Polyclonal Antibody

Paraformaldehyde-fixed, paraffin embedded (Rat 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-beta Catenin (Tyr333)) Polyclonal Antibody, Unconjugated at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Mouse testis); 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-beta Catenin (Tyr333)) Polyclonal Antibody, Unconjugated at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) instructionsand DAB staining.

Blank control (blue line): mouse brain (fixed with 2% paraformaldehyde for 10 min at room temperature, and then stained with Primary Antibody for 30 min at room temperature). Primary Antibody :Rabbit Anti-phospho-beta Catenin (Tyr333)antibody, Dilution: 1ug/10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-FTIC, Dilution: 1ug/test.

Blank control:293T. Primary Antibody (green line): Rabbit Anti-phospho-beta Catenin (Tyr333) antibody Dilution: 2ug/10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-FITC Dilution: 1ug/test. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature. 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.

Sample: Lane 1: Lung (Mouse) Lysate at 40 ug Lane 2: Cerebrum (Mouse) Lysate at 40 ug Lane 3: Cerebrum (Rat) Lysate at 40 ug Lane 4: Liver (Rat) Lysate at 40 ug Lane 5: Skin (Mouse) Lysate at 40 ug Lane 6: Skin (Rat) Lysate at 40 ug Primary: Anti-phospho-beta Catenin (Tyr333) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 92 kD Observed band size: 92 kD