Anti-trimethyl-Histone H3 (Lys36) Antibody, Trial Size

Histones are nuclear proteins that form octameric structures which bind DNA to form units of chromatin called nucleosomes. The family of histones—H2A, H2B, H3, and H4—are key players in gene regulation. They undergo a number of post-translational modifications (PTM) in response to various stimuli including phosphorylation on serine and threonine residues and methylation on lysine residues. PTMs produce configural changes in histone proteins that may induce nucleosome remodeling and expose or hide DNA sequences from transcriptional complexes. Methylation occurs on histone H3 at lysine 4, 9, 27, 36, and 79 residues. Previous studies have reported that methylation of H3 lysine 9 provides a binding site for heterochromatin adaptor proteins (HP1) which are involved in the formation of heterochromatin and gene repression. Phosphorylation of histones may also produce similar repressive effects depending on the cellular context. Often, it is the pattern of different modifications on several histone residues that alter chromatin, rather than modification of single residues.

Broad species reactivity expected due to similar sequence homology.

This antibody recognizes Histone H3 trimethylated at Lys36.

Epitope: Trimethylated Lys36

KLH-conjugated linear peptide corresponding to human Histone H3 trimethylated at Lys36.

Anti-trimethyl-Histone H3 (Lys36) Antibody, Trial Size, is a rabbit polyclonal antibody for detection of Histone H3 trimethylated at lysine 36 (H3K36me3). This purified Ab is specificity verified by dot blot (DB) and validated in WB, ICC. Small pack size is ideal for screening applications.

Dot Blot Specificity Analysis: Unmodified and various modified Histone peptides (see Table) were probed with Anti-trimethyl-Histone H3 (Lys36) (1:2,000 dilution).

Immunocytochemistry Analysis: A 1:500 dilution from a representative lot detected Histone H3 in A431 cells.

ChIP-Sequencing: Chromatin immunoprecipitation was performed using the Magna ChIP HiSens kit (cat#17-10460, 2 µg of anti-trimethyl-Histone H3 (Lys36) antibody(ABE435-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 eleven 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.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Histones

Evaluated by Western Blot in HeLa cell extract and recombinant Histone H3 protein.

Western Blot Analysis: A 1:500 dilution of this antibody detected Histone H3 on 10 µg of HeLa cell extract and recombinant Histone H3 protein.

~17 kDa observed. An uncharacterized band appears at ~52 kDa in some lysates.

Affinity purified

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

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

Control
HeLa cell extract and recombinant Histone H3 protein

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

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.