Smad1 Rabbit Polyclonal Antibody

Blank control: Hela. Primary Antibody (green line): Rabbit Anti-Smad1 antibody (orb11383), Dilution: 1 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, Dilution: 0.5 ug/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. 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.

Molt-4 cells were fixed with 4% PFA for 10 min at room temperature, permeabilized with 90% ice-cold methanol for 20 min at -20°C, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Smad1 Antibody (orb11383) at 1:500 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

Tissue/Cell: human lung carcinoma, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), 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-Smad1 Polyclonal Antibody, Unconjugated (orb11383) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.