Anti-dimethyl Histone H3 (Lys4) Antibody

产品说明

一般描述

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.

特异性

This antibody recognizes Histone H3 dimethylated at Lys4.
Broad species cross-reactivity expected based on sequence similarity.

免疫原

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

应用

Research Sub Category
Histones
Research Category
Epigenetics & Nuclear Function
Anti-dimethyl Histone H3 (Lys4) Antibody is a Rabbit Polyclonal Antibody for detection of Histone H3 dimethylated at lysine 4. This purified polyclonal antibody, also known as H3K4me2, has dot blot (DB) proven specificity & has been validated in WB, ICC, ChIP.
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 ofa representative lot of anti-dimethyl-Histone H3 (Lys4) antibody(ABE250), 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.

质量

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.

目标描述

~17 kDa observed

联系

Replaces: 04-791

外形

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

储存及稳定性

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

分析说明

Control
HeLa acid extract

免责声明

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.

基本信息

产品性质

安全信息