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Merck
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Key Documents

04-790

Sigma-Aldrich

Anti-dimethyl-Histone H3 (Lys4) Antibody, clone AW30, rabbit monoclonal

culture supernatant, clone AW30, Upstate®

Sinônimo(s):

H3K4me2, Histone H3 (di methyl K4)

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About This Item

Código UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.41

fonte biológica

rabbit

Nível de qualidade

forma do anticorpo

culture supernatant

tipo de produto de anticorpo

primary antibodies

clone

AW30, monoclonal

reatividade de espécies

chicken, human

fabricante/nome comercial

Upstate®

técnica(s)

ChIP: suitable (ChIP-seq)
dot blot: suitable
inhibition assay: suitable (peptide)
western blot: suitable

Isotipo

IgG

nº de adesão NCBI

nº de adesão UniProt

Condições de expedição

dry ice

modificação pós-traducional do alvo

dimethylation (Lys4)

Informações sobre genes

human ... H3C1(8350)

Descrição geral

Histones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. The four core histones, H2A, H2B, H3, and H4, assemble into an octamer (2 molecules of each). Subsequently, 146 base pairs of DNA are wrapped around the octamer, forming a nucleosome, the basic subunit of chromatin. Histones are modified post-translationally by the actions of enzymes in both the nucleus and cytoplasm. These modifications regulate DNA transcription, repair, recombination, and replication. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. These modifications occur predominantly on the N-terminal and C-terminal tails that extend beyond the nucleosome core particle. Di- and trimethylation of histone H3 at Lys4 correlates with transcriptional activity of many genes. Dimethylation of Histone H3 at Lys4 occurs at both active and inactive euchromatic regions but not in silent heterochromatic sites, whereas trimethylation at Lys4 is present exclusively at active genes.

Especificidade

Histone H3 dimethylated on lysine 4
The immunizing sequence is conserved from Tetrahymena to human, so broad species cross-reactivity is expected.

Imunogênio

KLH-conjugated, synthetic peptide containing the sequence …Tme2KQT… in which me2K corresponds to dimethyl-lysine at residue 4 of human histone H3.

Aplicação

Western Blot Analysis:
A 1:1000-1:2000 dilution of this lot detected dimethyl histone H3 (Lys4) in HeLa acid extracts.

Western Blot Analysis:
A 1:2,000 to 1:10,000 dilution of a previous lot detected methylated histone H3 in acid extracted proteins from HeLa cells. The antibody did not detect unmethylated recombinant Histone H3.

ChIP-seq Analysis:
Representative lot data. Chromatin immunoprecipitation was performed using the Magna ChIP HiSens kit (cat# 17-10460), Anti-dimethyl-Histone H3 (Lys4) antibody (2 µl cat# 04-790), 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 sixteen 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 04-790 and 05-1338 datasets showed 92 and 90% overlap with peaks identified in the ENCODE H3K4me2 BROAD Histone track for HeLa S3.

Dot Blot Analysis:
Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with Cat. No 04-790, Anti-dimethyl Histone H3 (Lys4), clone AW30 at 1:1000 dilution. Proteins were visualized using a Donkey anti-Rabbit IgG conjugated to HRP and a chemiluminescence detection system.

Peptide Inhibition Analysis:
2 μM of a histone H3 peptide containing dimethyl-lysine 4 abolished detection of histone H3 by a previous lot of antibody in immunoblot analysis of HeLa acid extracts. No signal reduction was observed with histone H3 peptides containing either monomethyl or trimethyl-lysine 4 modifications
Anti-dimethyl-Histone H3 (Lys4) Antibody, clone AW30 is a rabbit monoclonal antibody for detection of dimethyl-Histone H3 (Lys4) also known as H3K4me2, Histone H3 (di methyl K4) & has been validated in WB, ChIP, ChIP-seq, DB, PIA, Mplex.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Histones

Qualidade

routinely evaluated by immunoblot on acid-extracted proteins from HeLa cells; the antibody did not detect unmethylated recombinant Histone H3 (Catalog #14-494)

Descrição-alvo

~17kDa

Ligação

Replaces: 05-790

forma física

Cultured supernantant in 0.05% sodium azide

Armazenamento e estabilidade

Stable for 1 year at -20°C from date of receipt.
For maximum recovery of product, centrifuge the vial prior to removing the cap.

Nota de análise

Control
HeLa acid extracts

Informações legais

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

Exoneração de responsabilidade

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.

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Código de classe de armazenamento

12 - Non Combustible Liquids

Classe de risco de água (WGK)

WGK 1

Ponto de fulgor (°F)

Not applicable

Ponto de fulgor (°C)

Not applicable


Certificados de análise (COA)

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Structural basis of a histone H3 lysine 4 demethylase required for stem elongation in rice.
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