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17-615 Sigma-Aldrich

ChIPAb+ Acetyl-Histone H3 - ChIP Validated Antibody and Primer Set

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

Synonym: H3Ac, Histone H3 acetyl, Histone H3Ac

  •  eCl@ss 32160702

  •  NACRES NA.32

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Properties

Related Categories Antibodies, Antibodies for Epigenetics and Nuclear Signaling, ChIP Validated Antibodies, Primary Antibodies
biological source   rabbit
clone   polyclonal
species reactivity   Tetrahymena sp., human, human, mouse, mouse
mfr. no.   ChIPAb+
  Upstate®
application(s)   ChIP: suitable
  western blot: suitable
NCBI accession no.   NP_003484
UniProt accession no.   P68431
shipped in   dry ice

Description

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).

Specificity

Recognizes acetyl-histone H3

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

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

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).

Packaging

25 assays per set. ~5 μg per chromatin immunoprecipitation

Quality

Routinely evaluated by chromatin immunoprecipitation on HeLa nuclear extract.

Target description

17 kDa

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.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detection methods, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

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Peer-Reviewed Papers
15

References

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