Anti-Acetyl-Histone H3 (Lys9) Polyclonal Antibody is a Rabbit antibody targeting Acetyl-Histone H3 (Lys9). Anti-Acetyl-Histone H3 (Lys9) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Pig,Cow,Rabbit,Fruit Fly)

1. Tissue/cell: rat colon 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-Histone H3 (acetyl K9) Polyclonal Antibody, Unconjugated (TMAB-00035) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody (green line): Rabbit Anti-Histone H3 (acetyl K9) antibody (TMAB-00035), Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 μg/test.
3. Blank control: Molt4. Primary Antibody (green line): Rabbit Anti-Histone H3 (acetyl K9) antibody (TMAB-00035)
Dilution: 2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 μg/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.
4. Sample:
NIH/3T3 (Mouse) Cell Lysate at 30 μg
Hela (Human) Cell Lysate at 30 μg
MCF-7 (Human) Cell Lysate at 30 μg
U2os (Human) Cell Lysate at 30 μg
Primary: Anti-Histone H3 (acetyl K9) (TMAB-00035) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 15 kDa
Observed band size: 15 kDa

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthesised acetylpeptide: human Histone H3 around the acetylation site of K9

Human

ChIP antibodies,Methylated