E cadherin Rabbit Polyclonal Antibody

Blank control: Hepg2 (blue). Primary Antibody: Rabbit Anti-E cadherin antibody (orb500758, Green), 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-FITC (white blue), dilution: 1:200 in 1X PBS containing 0.5% BSA. Protocol, The cells were fixed with 2% paraformaldehyde for 10 min at 37°C. Primary antibody (orb500758, 1 µg/1x10^6 cells) were incubated for 30 min at room temperature, followed by 1X PBS containing 0.5% BSA + 1 0% goat serum (1 hour) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/FITC antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 40 min at room temperature. Acquisition of 20000 events was performed.

Tissue/Cell: mouse kidney tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-E-cadherin Polyclonal Antibody, Unconjugated (orb500758) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.