Phospho-beta Catenin (Ser33) Rabbit Polyclonal Antibody

Blank control (blue): RSC96 cells (fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice). Primary Antibody: Rabbit Anti-phospho-beta Catenin (Ser33) antibody (orb155825), Dilution: 1 µg 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-PE (white blue), Dilution: 1:200 in 1X PBS containing 0.5% BSA. Protocol, The cells were Antibody (orb155825, 1 µg/1x10^6 cells) were incubated for 30 min on the ice, followed by 1X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody of orb155825 at 1/200 dilution for 30 min on ice. Acquisition of 20000 events was performed.

Blank control: 293T. Primary Antibody (green line): Rabbit Anti-phospho-beta Catenin (Ser33) antibody (orb155825), Dilution: 2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-FITC, Dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min 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 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (human colon cancer), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (p-β Catenin) Polyclonal Antibody, Unconjugated (orb155825) at 1:500 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.