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05-1338

Sigma-Aldrich

Anti-Dimethyl Histone H3 (Lys4) Antibody, clone CMA303

clone CMA303, from mouse

Synonym(s):

H3K4me2, Histone H3 (di methyl K4), H3 histone family, member T, histone 3, H3, histone cluster 3, H3

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About This Item

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

biological source

mouse

Quality Level

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

CMA303, monoclonal

species reactivity

human, vertebrates

technique(s)

ChIP: suitable
ELISA: suitable
dot blot: suitable
immunocytochemistry: suitable
immunoprecipitation (IP): suitable
multiplexing: suitable
western blot: suitable

isotype

IgG1κ

NCBI accession no.

UniProt accession no.

shipped in

wet ice

target post-translational modification

dimethylation (Lys4)

Gene Information

human ... H3C1(8350)

General description

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. Histones are modified post-translationally; and these modifications regulate DNA transcription, repair, recombination, and replication. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. The modifications occur predominantly on the N-terminal and C-terminal tails that extend beyond the nucleosome core particle. Dimethyl-lysine 4 histone H3 (H3K4me2) is a transcription-activating chromatin mark, and dimethyl histone H3 Lys4 (H3K4me2) is depleted from regions with DNA methylation. Multipotential hematopoietic cells have a subset of genes that are differentially methylated (H3K4me2+/me3-). These genes are transcriptionally silent and highly enriched in lineage-specific hematopoietic genes, suggesting a role for H3K4 methylation in differentiation.

Specificity

Broad species cross-reactivity is expected, based on sequence homology.
This antibody specifically recognizes Histone H3 dimethylated at Lys4. The antibody binding specificity allows for phosphorylation of Thr3 and/or modifications of Lys91.

Immunogen

Epitope: Dimethylated Lys4
Synthetic peptide corresponding to amino acids 1-12 of human Histone H3, dimethylated on Lys4, conjugated to KLH.

Application

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG, or Anti-dimethyl-Histone H3 (Lys4) antibody and the Magna ChIP G (Cat. # 17-611) Kit. Successful immunoprecipitation of dimethyl-histone H3 (Lys4) associated DNA fragments was verified by qPCR using ChIP Primers GAPDH Coding.
Please refer to the EZ-Magna G ChIP (Cat. # 17-409) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

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 05-1338, Anti-dimethyl H3 (Lys4), clone CMA303 at 2.0 µg/mL (1:500 dilution). Proteins were visualized using a Donkey anti-mouse IgG conjugated to HRP and a chemiluminescence detection system.

ChIP-seq Analysis:
Chromatin immunoprecipitation was performed using the Magna ChIP HiSens kit (cat# 17-10460), 2 µg of Anti-dimethyl-Histone H3 (Lys4) antibody (cat# 05-1338), 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.

Immunocytochemistry:
This antibody has been shown by an outside laboratory to be suitable for immunocytochemistry.

Immunoprecipitation:
This antibody has been shown by an outside laboratory to be suitable for immunoprecipitation.

ELISA:
This antibody has been shown by an outside laboratory to be suitable for ELISA

Multiplexing:
This antibody specifically recognizes histone H3 dimethylated on Lys4 by Luminex assay.
Anti-Dimethyl Histone H3 (Lys4) Antibody, clone CMA303 is a mouse monoclonal antibody for detection of Dimethyl Histone H3 (Lys4) also known as H3K4me2, Histone H3 (di methyl K4). Demonstrated performance in ChIP, DB, ELISA, ICC, IP, WB, Mplex.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Histones

Quality

Routinely evaluated by Western Blot on HeLa acid extract.

Western Blot Analysis: 2 μg/mL (1:500) dilution of this antibody detected dimethyl Histone H3 (Lys4) on 10 μg of HeLa acid extract.

Target description

approx. 17 kDa

Physical form

Format: Purified
Protein G Purified
Purified mouse monoclonal IgG1κ in PBS with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt. For maximum recovery of product, centrifuge the vial prior to removing the cap. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Analysis Note

Control
HeLa Acid extract lysate

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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.

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Lihong Shi et al.
Nature medicine, 19(3), 291-294 (2013-02-19)
Enhanced fetal γ-globin synthesis alleviates symptoms of β-globinopathies such as sickle cell disease and β-thalassemia, but current γ-globin-inducing drugs offer limited beneficial effects. We show here that lysine-specific demethylase 1 (LSD1) inhibition by RNAi in human erythroid cells or by
Laura S Shankman et al.
Nature medicine, 21(6), 628-637 (2015-05-20)
Previous studies investigating the role of smooth muscle cells (SMCs) and macrophages in the pathogenesis of atherosclerosis have provided controversial results owing to the use of unreliable methods for clearly identifying each of these cell types. Here, using Myh11-CreER(T2) ROSA
Mike V Van et al.
Genetics, 204(3), 999-1013 (2016-09-09)
Meiotic recombination establishes connections between homologous chromosomes to promote segregation. Hemizygous regions of sex chromosomes have no homologous chromosome to recombine with, yet must be transmitted through meiosis. An extreme case of hemizygosity exists in the genus Caenorhabditis, where males
DNA methylation dictates histone H3K4 methylation.
Okitsu, Cindy Yen and Hsieh, Chih-Lin
Molecular and cellular biology, 27, 2746-2757 (2007)
Takashi Miwa et al.
Genes to cells : devoted to molecular & cellular mechanisms, 24(5), 377-389 (2019-04-01)
In Caenorhabditis elegans, germline cells remain transcriptionally silenced during embryogenesis. The transcriptional silencing is achieved by two different mechanisms: One is the inhibition of RNA polymerase II in P2-P4 cells at the establishment stage, and another is chromatin-based silencing in

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