ENO3 Rabbit Polyclonal Antibody

Blank control: Hela. Primary Antibody (green line): Rabbit Anti-ENO3 antibody (orb5157), dilution: 2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-PE, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 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 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.

Sample: Lane 1: Mouse Liver tissue lysates, Lane 2: Mouse Cerebrum tissue lysates, Lane 3: Rat Pancreas tissue lysates, Lane 4: Rat Liver tissue lysates, Lane 5: Rat Cerebrum tissue lysates, Lane 6: Human HeLa cell lysates, Lane 7: Human HepG2 cell lysates, Lane 8: Human A549 cell lysates, Lane 9: Human Jurkat cell lysates, Primary: Anti-ENO3 (orb5157) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 47 kDa, Observed band size: 47 kDa.

Tissue/Cell: rat brain 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-ENO3 Polyclonal Antibody, Unconjugated (orb5157) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.