Description
General description
17 kDa
All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ Acetyl-Histone H3 set contains purified rabbit polyclonal antibody and the normal rabbit IgG antibody, which can be used to demonstrate that the acetyl-histone H3 antibody is capable of precipitating acetyl-histone H3 associated chromatin. The qPCR primers included flank the human GAPDH promoter and produce a 166 base pair PCR product.
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. Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
Immunogen
The acetyl-histone H3 antibody is made against a peptide corresponding to amino acids 1-20 of Tetrahymena histone H3 ARTKQTAR[K*]STGG[K*]APRKQL(C) where K* is acetylated)
Application
Acetyl-Histone H3 ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of H3Ac.
Western Blot Analysis:
Acid-extracted proteins from HeLa cells treated with 5 mM sodium butyrate for 24 hours (Lane 2) and untreated HeLa cells (Lane 1) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-acetyl-Histone H3 (0.01 μg/mL).
Proteins were visualized using a goat-rabbit secondary antibody conjugated to HRP and a chemilumnescence detection system (Please see figures).
Biochem/physiol Actions
Recognizes acetyl-histone H3
Tetrahymena predicted to cross-react based upon sequence of antibody generating peptide.
Packaging
25 assays per set. ~5 μg per chromatin immunoprecipitation
Physical form
Anti-Acetyl-Histone H3 (rabbit polyclonal IgG). One vial containing 125 ug of protein A purified antibody in 125 μL of storage buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide. Store at -20°C.
Normal Rabbit IgG. One vial containing 125 ug of normal rabbit IgG in 125 uL volume. Store at -20°C.
Control Primers. One vial containing 75 μL of 5 μM of each primer specific for for human GAPDH. Store at -20°C.
FOR: TAC TAG CGG TTT TAC GGG CG
REV: TCG AAC AGG AGG AGC AGA GAG
CGA
Format: Purified
Analysis Note
Control
Included negative control antibody rabbit IgG and control primers specific for human GAPDH promoter.
Routinely evaluated by chromatin immunoprecipitation on HeLa nuclear extract.
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
