Application

ChIP-seq Analysis:
Representative lot data. Chromatin immunoprecipitation was performed using the Magna ChIP™ HiSens kit (cat# 17-10460), 3 µL of Anti-trimethyl-Histone H3 (Lys4) antibody (cat# 05-745R-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 eighteen 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 05-745R and 07-473 datasets showed 99% overlap with peaks identified in the ENCODE H3K4me3 BROAD Histone track for HeLa S3.

Research Sub CategoryHistones

Research CategoryEpigenetics & Nuclear Function

Use Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, Recombinant, Trial Size (rabbit monoclonal antibody) proven in WB, ChIP, ChIP-seq, ELISA, PIA, Mplex, DB to detect trimethyl-Histone H3 (Lys4) also known as H3K4me3.

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

Recombinant version of Cat. # 05-745, clone MC315

Immunogen

BSA-conjugated, synthetic peptide containing the sequence R30;RTme3KQTR30; in which me3K corresponds to trimethyl-lysine 4 of human histone H3.

Legal Information

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

Linkage

Replaces: 05-745

Physical form

Protein A purified, recombinant antibody in 60 mM Tris, 90 mM NaCl, 0.03% NaN3, 60 mM Glycine, pH 7.5, 40% Glycerol.

Format: Purified

Quality

Routinely evaluated by immunoblot on HeLa nuclear extract

Specificity

Recognizes Histone H3 containing trimethyl-lysine 4 and, to a lesser extent, dimethyl-lysine 4. See certificate of analysis for further details.

Broad species cross-reactivity expected due to sequence homology

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol-containing solutions to become frozen during storage.

Target description

17 kDa