Anti-Acetyl-Histone H4 (Lys8) Polyclonal Antibody 3 is a Rabbit antibody targeting Acetyl-Histone H4 (Lys8). Anti-Acetyl-Histone H4 (Lys8) Polyclonal Antibody 3 can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Dog,Pig,Cow,Horse,Rabbit)

1. 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 (TMAB-07137) 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.
2. 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 (TMAB-07137) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. 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 (TMAB-07137) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
4. 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) (TMAB-07137) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 11 kD
Observed band size: 11 kD
5. Blank control (black line): NIH/3T3.
Primary Antibody (green line): Rabbit Anti-Histone H4 (Acetyl K8) antibody (TMAB-07137)
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/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℃, 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 20,000 events was performed.

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

Polyclonal

KLH conjugated Synthesised acetylpeptide: human Histone H4 around the acetylation site of Lys8

Human

H4C1

Histone H4

Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.