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Key Documents

04-808

Sigma-Aldrich

Anti-dimethyl-Histone H3 (Arg2), clone 20.2 Antibody, rabbit monoclonal

culture supernatant, clone 20.2, Upstate®

Synonym(s):

H3R2me2, Histone H3 (di methyl R2)

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About This Item

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

biological source

rabbit

Quality Level

antibody form

culture supernatant

antibody product type

primary antibodies

clone

20.2, monoclonal

species reactivity

human, vertebrates

manufacturer/tradename

Upstate®

technique(s)

dot blot: suitable
inhibition assay: suitable (peptide)
multiplexing: suitable
western blot: suitable

isotype

IgG

NCBI accession no.

UniProt accession no.

shipped in

dry ice

target post-translational modification

dimethylation (Arg2)

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, the basic subunit of chromatin. Histones are modified post-translationally by the actions of enzymes in both the nucleus and cytoplasm. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. These modifications can alter local chromatin architecture, or recruit trans-acting factors that recognize specific histone modifications (the "histone code" hypothesis). The modifications occur predominantly on the N-terminal and C-terminal tails that extend beyond the nucleosome core particle. Histone H3 Arg2 methylation is a widespread silencing modification that inhibits histone H3 Lys4 trimethylation.

Specificity

Recognizes dimethyl-arginine at residue 2 of histone H3, Mr 17kDa. Additional unidentified bands above 50 kDa are detected in some samples.
The immunizing sequence is highly conserved evolutionarily, so broad cross-reactivity is expected.

Immunogen

Epitope: Arg2 of Histone H3
KLH-conjugated, synthetic peptide containing the sequence …Ame2RTKQ…, in which me2R corresponds to dimethyl-arginine 2 of human histone H3.

Application

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. 04-808, Anti-dimethyl H3 (Arg2) at 1ug/mL (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 histone H3 peptide containing dimethyl-Arg2 abolished detection of histone H3 by a previous lot of anti-dimethyl-Histone H3(Arg2) (1:4,000 dilution) in immunoblots of HeLa acid extracts. Some competition of signal was also seen with unmodified peptide containing Arg2.

Peptide Dot Blot Analysis:
A 1:4,000-1:16,000 dilution of a previous lot only detected histone H3 peptide containing dimethyl-Arg2. Peptides containing unmodi-fied Arg2, dimethyl-Arg17 or dimethyl-Arg26 were not detected.

Beadlyte Histone Peptide Assay:
1:500-1:48,000 dilutions of a previous lot detected histone H3 peptide containing dimethyl-Arg2. Cross-reactivity to peptides containing unmodified Arg2 or dimethylArg 17 or 26 was not seen.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Histones
Use Anti-dimethyl-Histone H3 (Arg2), clone 20.2 Antibody (rabbit monoclonal antibody) validated in DB, Mplex, PIA, WB to detect dimethyl-Histone H3 (Arg2) also known as H3R2me2, Histone H3 (di methyl R2).

Quality

Routinely evaluated by immunoblot on acid-extracted proteins from HeLa cells.

Immunoblot Analysis: A 1:500 (lane 1) and 1:1000 (lane 2) dilution of this lot detected methylated Histone H3 in acid-extracted proteins from HeLa cells.

Target description

17kDa

Linkage

Replaces: 05-808

Physical form

Cultured supernantant in 0.05% sodium azide

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Analysis Note

Control
HeLa cell acid 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.

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

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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Yudao Shen et al.
Journal of medicinal chemistry, 59(19), 9124-9139 (2016-09-02)
Well-characterized selective inhibitors of protein arginine methyltransferases (PRMTs) are invaluable chemical tools for testing biological and therapeutic hypotheses. Based on 4, a fragment-like inhibitor of type I PRMTs, we conducted structure-activity relationship (SAR) studies and explored three regions of this
Xin-Xi Zhao et al.
The Journal of biological chemistry, 291(1), 402-412 (2015-10-22)
Histone lysine methylation is important in early zebrafish development; however, the role of histone arginine methylation in this process remains unclear. H3R2me2a, generated by protein arginine methyltransferase 6 (Prmt6), is a repressive mark. To explore the role of Prmt6 and
Histone H3 Arg2 methylation provides alternative directions for COMPASS.
Klose, Robert J and Zhang, Yi
Nature Structural and Molecular Biology, 14, 1058-1060 (2007)
Yudao Shen et al.
Journal of medicinal chemistry, 64(7), 3697-3706 (2021-02-17)
Protein arginine methyltransferase 6 (PRMT6) catalyzes monomethylation and asymmetric dimethylation of arginine residues in various proteins, plays important roles in biological processes, and is associated with multiple cancers. To date, a highly selective PRMT6 inhibitor has not been reported. Here
Daniel J Slade et al.
Biopolymers, 101(2), 133-143 (2013-04-12)
Post-translational modifications (PTMs) of protein embedded arginines are increasingly being recognized as playing an important role in both prokaryotic and eukaryotic biology, and it is now clear that these PTMs modulate a number of cellular processes including DNA binding, gene

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