Analysis Note

ControlSodium butyrate untreated and treated HeLa acid extract

Application

Research Sub CategoryHistones

Anti-acetyl Histone H3 (Lys9) Antibody, Trial Size is a rabbit polyclonal for detection of Histone H3 acetylated at lysine 9. Also known as Anti-H3K9ac, this antibody has dot blot (DB) proven specificity and has been validated in WB, ChIP, Mplex. Small pack size is ideal for screening applications

Research CategoryEpigenetics & Nuclear Function

Chromatin Immunoprecipitation Analysis (ChIP):
A representative lot of this antibody immunoprecipitated chromatin from Histone H3 acetylated on Lys9.

ChIP-Sequencing:
Chromatin immunoprecipitation was performed using the Magna ChIP™ HiSens kit (cat# 17-10460), 2 μg of a representative lot of anti-acetyl-Histone H3 (Lys9) antibody (ABE18-S), 20μL Protein A/G beads, and 1e6 crosslinked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using standard protocols with Illumina barcoded adapters, and analyzed on Illumina HiSeq instrument. An excess of thirteen million reads from FastQ files were mapped using Bowtie(http://bowtie-bio.sourceforge.net/manual.shtml) following TagDust (http://genome.gsc.riken.jp/osc/english/dataresource/) tag removal. Peaks were identified using MACS (http://luelab.dfci.harvard.edu/MACS/), with peaks and reads visualized as a custom track in UCSC Genome Browser (http://genome.ucsc.edu) from BigWig and BED files.

Dot Blot Analysis:
A representative lot pf this antibody detected Histone H3 in modified and non modified peptides of acetylated and non-acetylated Histone H3 Lys9.

Luminex Analysis:
A representative lot of this antibody specifically recognized Histone H3 acetylated on Lys9 by Luminex™ assay.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the ′beads on a string′ structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine. Acetylation of histone H3 occurs at several different lysine positions in the histone tail and is performed by a family of enzymes known as Histone Acetyl Transferases (HATs). Acetylation at lysine 9 is believed to play a role in histone deposition and chromatin assembly.

Immunogen

KLH-conjugated linear peptide corresponding to human Histone H3 acetylated at Lys9.

Epitope: Acetylated Lys9

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Affinity purified

Purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Quality

Evaluated by Western Blot in sodium butyrate untreated and treated HeLa acid extract.

Western Blot Analysis: 0.05 µg/mL of this antibody detected Histone H3 on 10 µg of sodium butyrate untreated and treated HeLa acid extract.

Specificity

Broad species cross-reactivity is expected.

This antibody recognizes Histone H3 when acetylated at Lys9.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Target description

~17 kDa observed