Application

Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either Normal Rabbit Serum , or 2 µL of Anti-Acetyl-Histone H3 (Lys18)and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Acetyl-Histone H3 (Lys18) associated DNA fragments was verified by qPCR using Control Primers specific for the human GAPDH promoter region as a positive locus, and β-globin primers as a negative locus. Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

ChIP-Sequencing:
Representative lot data. Chromatin immunoprecipitation was performed using the Magna ChIP™ HiSens kit (cat# 17-10460), 2 μg of anti-acetyl-Histone H3 (Lys18) antibody (cat# 07-354), 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 ten 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.

Western Blot Analysis:
Representative lot data.
Recombinant histone H3 (lane 1, Catalog # 14-494) and acid extracts from sodium butyrate treated (lane 2) and untreated (lane 3) HeLa cells (Catalog # 17-305) were probed with anti acetyl- Histone H3 (Lys18) (1:10,000 dilution).
Arrow indicates acetyl histone H3 (Lys18) (17 kDa).

Dot Blot:
Representative lot data.
40 ng and 4ng amounts of histone peptides with various modifications (see table 1) were transferred to PVDF membrane and probed with Anti-Acetyl-Histone H3 (Lys18) antibody (1:5000 dilution). Proteins were visualized using a goat anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system. Image from a 60 second exposure is shown.

Use Anti-acetyl-Histone H3 (Lys18) Antibody (Rabbit Polyclonal Antibody) validated in ChIP, WB to detect acetyl-Histone H3 (Lys18) also known as H3K18Ac, Histone H3 (acetyl K18).

General description

Histone H3 is one of the 5 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>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).

Immunogen

KLH-conjugated, synthetic peptide (GKAPRAcKQLASK-C) corresponding to amino acids 13-23 of yeast Histone H3 acetylated on lysine 18with a C-terminal cysteine added for conjugation purposes

Legal Information

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

Linkage

Replaces: 04-1107

Other Notes

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

Physical form

100 µLof rabbit serum containing 0.05% sodium azide and 30% glycerol.

Quality

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

Specificity

Recognizes Histone H3 acetylated on lysine 18.

Target description

17 kDa