Histone H4 (Acetyl K8) Rabbit Polyclonal Antibody

Blank control (black line): NIH/3T3. Primary Antibody (green line): Rabbit Anti-Histone H4 (Acetyl K8) antibody (orb5662), 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.

Hela cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (Histone H4 (Acetyl K8)) polyclonal Antibody, Unconjugated (orb5662) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Paraformaldehyde-fixed, paraffin embedded (Rat liver), 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 (Histone H4 (Acetyl K8)) Polyclonal Antibody, Unconjugated (orb5662) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat skin), 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 (Histone H4 (Acetyl K8)) Polyclonal Antibody, Unconjugated (orb5662) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

Sample: Lane 1: Human Hela cell lysates, Lane 2: Human Jurkat cell lysates, Lane 3: Human HL60 cell lysates, Lane 4: Human MCF-7 cell lysates, Primary: Anti-Histone H4 (Acetyl K8) (orb5662) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 11 kD, Observed band size: 11 kD.