Anti-dimethyl Histone H3 (Lys4) Antibody, Trial Size (C15-1317-038)

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. Dimethylation of histone H3 at Lys4 correlates with transcriptional activity of many genes.

Specificity

Broad species cross-reactivity expected based on sequence similarity.

This antibody recognizes Histone H3 dimethylated at Lys4.

Immunogen

Epitope: Dimethylated Lys4

KLH-conjugated linear peptide corresponding Histone H3 methylated at Lys4.

Application

Anti-dimethyl Histone H3 (Lys4) Antibody, Trial Size is a Rabbit Polyclonal for detection of Histone H3 dimethylated at lysine 4. Also known as H3K4me2, this Ab has dot blot (DB) proven specificity & has been validated in WB, ICC, ChIP. Small pack size is ideal for screening applications.

Immunocytochemistry Analysis:
1:500 dilution from a representative lot detected Histone H3 in NIH/3T3 and A431 cells.

Dot Blot Analysis:
1:1,000 dilution from a representative lot specifically detected Histone H3 in a Lys4 dimethhyl modified histone H3 peptide while not detecting potential cross-reacting peptides corresponding to other modified and non-modified histones.

Chromatin Immunoprecipitation Analysis:
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 4 µg of either Normal IgG (Part No. 12-370), or 4 µL of Anti-Dimethyl Histone H3 (Lys4) (Part No. ABE250) and the Magna ChIP A/G Kit (Cat. # 17-10085). Successful immunoprecipitation of Dimethyl-Histone H3 (Lys4) associated DNA fragments was verified by qPCR using primers specific for the human GAPDH coding region.
Please refer to the EZ-Magna ChIP A/G (Cat. # 17-10085) protocol for experimental details.

ChIP-Sequencing
Chromatin immunoprecipitation was performed using the Magna ChIP HiSens kit (cat# 17-10460), 5 µg of a representative lot of anti-dimethyl-Histone H3 (Lys4) antibody(ABE250-S), 20 µL Protein A/G beads ,and 5e6 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 twelve 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

Quality

Evaluated by Western Blot using HeLa acid extract and recombinant histone proteins.

Western Blot Analysis: 1:1,000 dilution of this antibody detected Histone H3 on 10 µg of HeLa acid extract.

Target description

~17 kDa observed

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.

Storage and Stability

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

Analysis Note

Control
HeLa acid extract

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.

biological source: rabbit. Quality Level: 100. antibody form: affinity isolated antibody. antibody product type: primary antibodies. clone: polyclonal. purified by: affinity chromatography. species reactivity: rat, chicken, human, mouse. technique(s): ChIP: suitable (ChIP-seq), dot blot: suitable, immunocytochemistry: suitable, western blot: suitable. NCBI accession no.: NP_003484.1. UniProt accession no.: Q16695. shipped in: wet ice. target post-translational modification: dimethylation (Lys4). Gene Information: human ... HIST1H3F(8968). Storage Class Code: 12 - Non Combustible Liquids. WGK: WGK 1. Flash Point(F): Not applicable. Flash Point(C): Not applicable.