DATASHEET
Host:
Rabbit
Target Protein:
H4K8ac
Clonality:
Polyclonal
Isotype:
Not Determined
Entrez Gene:
121504
Swiss Prot:
P62805
Source:
Polyclonal antibody raised in rabbit against histone H4 containing the acetylated lysine 8 (H4K8ac), using a KLH-conjugated synthetic peptide
Purification:
Whole antiserum from rabbit containing 0.05% azide
Storage Buffer:
0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage:
Store at -20°C for 12 months.
Background:
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.
VALIDATION IMAGES
To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against H4K8ac Polyclonal Antibody (bs-53063R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:17,500.
A Dot Blot analysis was performed to test the cross-reactivity of
H4K8ac Polyclonal Antibody (bs-53063R) with peptides containing other modifications of histone H4 and H3 and an unmodified histone H4 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the modification of interest.
Histone extracts of HeLa cells (15 μg) were analyzed by Western blot using the antibody against H4K8ac (bs-53063R) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. Lane 2 shows the result of the Western analysis with the antibody; lane 1 shows the same analysis after incubation of the antibody with 750 pmol blocking peptide for 1 hour at room temperature.