Phospho-HDAC3 (Ser424) Rabbit Polyclonal Antibody

Blank control (black line): Molt4. Primary Antibody (green line): Rabbit Anti-Phospho-HDAC3 (Ser424) antibody (orb5754), dilution: 1 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488, dilution: 0.5 ug/Test. Isotype control (orange line): Normal Rabbit IgG, 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.

Blank control (black line): NIH/3T3. Primary Antibody (green line): Rabbit Anti-Phospho-HDAC3 (Ser424) antibody (orb5754), dilution: 1 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488, dilution: 0.5 ug/Test. Isotype control (orange line): Normal Rabbit IgG, 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.

Sample: Lane 1: Mouse NIH/3T3 cell lysates, Lane 2: Mouse Testis tissue lysates, Lane 3: Mouse Cerebrum tissue lysates, Lane 4: Rat Cerebrum tissue lysates, Primary: Anti-Phospho-HDAC3 (Ser424) (orb5754) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 47 kDa, Observed band size: 50 kDa.

Sample: MCF-7 (Human) Cell Lysate at 30 ug, NIH/3T3 (Mouse) Cell Lysate at 30 ug, Hela (Human) Cell Lysate at 30 ug, Primary: Anti-Phospho-HDAC3 (Ser424) (orb5754) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 50 kD, Observed band size: 50 kD.

Tissue/Cell: Rat rectum tissue, 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-Phospho-HDAC3, Unconjugated (orb5754) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.