07-473

Sigma-Aldrich

Anti-trimethyl-Histone H3 (Lys4) Antibody

Upstate®, from rabbit

Synonym(s):
H3K4me3, Histone H3 (tri methyl K4), H3 histone family, member T, histone 3, H3, histone cluster 3, H3
eCl@ss:
32160702
NACRES:
NA.41

biological source

rabbit

Quality Level

antibody form

affinity isolated antibody

antibody product type

primary antibodies

clone

polyclonal

purified by

affinity chromatography

species reactivity

human, vertebrates

manufacturer/tradename

Upstate®

application(s)

ChIP: suitable (ChIP-seq)
dot blot: suitable
western blot: suitable

NCBI accession no.

UniProt accession no.

shipped in

wet ice

General description

Histones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. Histone modifications regulate DNA transcription, repair, recombination, and replication. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which DNA is wrapped in repeating units, called nucleosomes, which limits DNA accessibility to the cellular machineries, which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Histone H3 contains a main globular domain and a long N-terminal tail and 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. H3K4me3 modifications are reported to occur consistently at transcription start sites and H3K4me3 domain is associated with higher transcription activity and cell identity in pre-implantation development and in the process of deriving embryonic stem cells from the inner cell mass and trophoblast stem cells from the trophectoderm. (Ref.: Liu, X., et al. (2016). Nature 537(7621); 558-562).

Specificity

This antibody only detects Histone H3 when trimethylated on Lysine 4.
Expected to react with most other species based on extreme conserved homology among species.

Immunogen

Epitope: Trimethylated (Lys4)
Synthetic peptide corresponding to residues surrounding and including trimethlyated Lys 4 of Histone H3.

Application

Research Sub Category
Histones
Research Category
Epigenetics & Nuclear Function
ChIP Analysis:
An independent lab has shown that this antibody performs in chromatin immunoprecipitation (ChIP).
Dot Blot Analysis: A 1:10,000 dilution from a representative lot detected trimethyl-Histone H3 (Lys 4 ) in an Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and an Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665).
ChIP-seq Analysis:
Chromatin immunoprecipitation was performed using the Magna ChIP HiSens kit (cat# 17-10460), 1 µL Anti-trimethyl-Histone H3 (Lys4) antibody (cat# 07-473) , 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 eighteen 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 07-473 dataset showed 99% overlap with peaks identified in the ENCODE H3K4me3 BROAD Histone track for HeLa S3.
Anti-trimethyl-Histone H3 (Lys4) Antibody is a rabbit polyclonal antibody for detection of Histone H3 trimethylated at lysine 4. Also known as Anti-H3K4me3, this highly specific and well published antibody has been validated in ChIP, DB, WB, PIA, ChIP-seq.

Quality

Evaluated by Western Blotting on HeLa acid extracted nuclear preps (positive) and recombinant Histone H3 (negative).

Western Blotting Analysis:
1:5,000 – 1:10,000 dilution of this antibody was used to detect trimethylated-Histone H3 (Lys4) in HeLa nuclear acid extracts.

Target description

~17 kDa observed; 15.51 kDa calculated. Uncharacterized bands may be observed in some lysate(s).

Linkage

Replaces: 04-791

Physical form

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

Storage and Stability

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

Analysis Note

Control
HeLa nuclear extracts

Legal Information

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

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.

Certificate of Analysis

Certificate of Origin

HISTONE MONOUBIQUITINATION1 interacts with a subunit of the mediator complex and regulates defense against necrotrophic fungal pathogens in Arabidopsis.
Dhawan, R; Luo, H; Foerster, AM; Abuqamar, S; Du, HN; Briggs, SD; Mittelsten Scheid, O; Mengiste, T
Plant Cell null
Immunoassay for human serum hepcidin.
Tomas Ganz, Gordana Olbina, Domenico Girelli, Elizabeta Nemeth, Mark Westerman
Blood null
Attenuation of brassinosteroid signaling enhances FLC expression and delays flowering.
Domagalska, MA; Schomburg, FM; Amasino, RM; Vierstra, RD; Nagy, F; Davis, SJ
Development null
Maintenance of long-range DNA interactions after inhibition of ongoing RNA polymerase II transcription.
Palstra, RJ; Simonis, M; Klous, P; Brasset, E; Eijkelkamp, B; de Laat, W
Testing null
Whole-genome analysis of histone H3 lysine 4 and lysine 27 methylation in human embryonic stem cells.
Pan, G; Tian, S; Nie, J; Yang, C; Ruotti, V; Wei, H; Jonsdottir, GA; Stewart, R; Thomson, JA
Cell Stem Cell null

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service

Social Media

LinkedIn icon
Twitter icon
Facebook Icon
Instagram Icon

MilliporeSigma

Research. Development. Production.

We are a leading supplier to the global Life Science industry with solutions and services for research, biotechnology development and production, and pharmaceutical drug therapy development and production.

© 2021 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.

Reproduction of any materials from the site is strictly forbidden without permission.