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Analysis Note

ControlIncluded negative control antibody rabbit IgG and control primers specific for human GAPDH promoter.

Application

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).

Acetyl-Histone H3 ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of H3Ac.

General description

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)

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

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

Quality

Routinely evaluated by chromatin immunoprecipitation on HeLa nuclear extract.

Specificity

Recognizes acetyl-histone H3

Tetrahymena predicted to cross-react based upon sequence of antibody generating peptide.

Target description

17 kDa