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Merck
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重要文件

04-790

Sigma-Aldrich

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

culture supernatant, clone AW30, Upstate®

同義詞:

H3K4me2, Histone H3 (di methyl K4)

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About This Item

分類程式碼代碼:
12352203
eCl@ss:
32160702
NACRES:
NA.41

生物源

rabbit

品質等級

抗體表格

culture supernatant

抗體產品種類

primary antibodies

無性繁殖

AW30, monoclonal

物種活性

chicken, human

製造商/商標名

Upstate®

技術

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

同型

IgG

NCBI登錄號

UniProt登錄號

運輸包裝

dry ice

目標翻譯後修改

dimethylation (Lys4)

基因資訊

human ... H3C1(8350)

一般說明

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.

特異性

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

免疫原

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

應用

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

品質

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

標靶描述

~17kDa

聯結

Replaces: 05-790

外觀

Cultured supernantant in 0.05% sodium azide

儲存和穩定性

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

分析報告

Control
HeLa acid extracts

法律資訊

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

免責聲明

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.

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儲存類別代碼

12 - Non Combustible Liquids

水污染物質分類(WGK)

WGK 1

閃點(°F)

Not applicable

閃點(°C)

Not applicable


分析證明 (COA)

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Barbara Meller et al.
Frontiers in plant science, 9, 1228-1228 (2018-09-21)
In this paper we analyzed β-aminobutyric acid (BABA)-primed epigenetic adjustment of potato cv. "Sarpo Mira" to Phytophthora infestans. The first stress-free generation of the potato genotype obtained from BABA-primed parent plants via tubers and seeds showed pronounced resistance to the
A role for Set1/MLL-related components in epigenetic regulation of the Caenorhabditis elegans germ line.
Li, T; Kelly, WG
PLoS Genetics null
Structural basis of a histone H3 lysine 4 demethylase required for stem elongation in rice.
Chen, Q; Chen, X; Wang, Q; Zhang, F; Lou, Z; Zhang, Q; Zhou, DX
PLoS Genetics null
X Zhao et al.
Oncogene, 35(27), 3565-3576 (2015-11-10)
Previous studies have indicated the important roles of MYCN in tumorigenesis and progression of neuroblastoma (NB), the most common extracranial solid tumor derived from neural crest in childhood. However, the regulatory mechanisms of MYCN expression in NB still remain largely
Rachel Bayley et al.
Molecular cell, 82(10), 1924-1939 (2022-04-20)
The 53BP1-RIF1-shieldin pathway maintains genome stability by suppressing nucleolytic degradation of DNA ends at double-strand breaks (DSBs). Although RIF1 interacts with damaged chromatin via phospho-53BP1 and facilitates recruitment of the shieldin complex to DSBs, it is unclear whether other regulatory

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