Analysis Note

ControlAcid extracted proteins from HeLa cells treated with sodium butyrate

Application

Anti-acetyl-Histone H3 (Lys27) Antibody is a Rabbit Polyclonal Antibody for detection of Histone H3 acetylated at lysine 27 (H3K27ac). This antibody has dot blot verified specificity and has been validated in ChIP, WB & Mplex.

Research Sub CategoryHistones

Research CategoryEpigenetics & Nuclear Function

ChIP-seq Analysis: Chromatin immunoprecipitation was performed using the Magna ChIP™ HiSens kit (cat# 17-10460), 2 µl of anti-acetyl-Histone H3 (Lys27) antibody (cat# 07-360-S), 20 µL 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 fourteen 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. The highest 25% of peaks identified in the 07-360 dataset showed 96% overlap with peaks identified in the ENCODE H3K27Ac BROAD Histone track for HeLa S3.

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. r> r>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.

Immunogen

Synthetic peptide corresponding to the sequence AR(acK)SA, in which acK corresponds to acetyl lysine 27 of yeast histone H3.

Epitope: Lys27

Legal Information

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

Other Notes

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

Physical form

Immunoaffinity Purified immunoglobulin in 0.2M Tris-glycine, pH 7.4, 0.15M NaCl, 0.05% sodium azide, and 5mg/ml BSA before the addition of glycerol to 30%

Unpurified

Quality

routinely evaluated by immunoblot on acid extracts from sodium butyrate treated HeLa cells

Specificity

Recognizes Histone H3 acetylated on lysine 27.

Storage and Stability

Maintain for 2 years at -20°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Target description

17 kDa