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05-1249

Sigma-Aldrich

Anti-Dimethyl Histone H3 (Lys9) Antibody, clone CMA307

clone CMA307, 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

CMA307, monoclonal

species reactivity

vertebrates, human

technique(s)

ChIP: suitable
ELISA: 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 (Lys9)

Gene Information

human ... H3C1(8350)

General description

Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the ′beads on a string′ structure. Histone proteins are highly post-translationally modified, and Histone H3 is the most extensively modified of the five histones. Histone H3 sequence variants and variable modification states are thought to play a role in the dynamic and long term regulation of genes. Di- and tri-methylation of histone H3 are gene silencing marks, while removal of these marks by the demethylase genes Jmjd1a and Jmjd2c leads to gene activation.

Specificity

Broad species cross-reactivity is expected, based on sequence homology.
This antibody recognizes Histone H3 dimethylated at Lys9. Phosphorylation of Ser10 interferes with antibody binding to dimethyl Lys9.

Immunogen

Epitope: dimethyl Lys9
Synthetic peptide corresponding to amino acids 1-19 of human Histone H3, dimethylated on Lys9, 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 (Lys9) antibody, clone CMA307 and the Magna ChIP G (Cat. # 17-611) Kit. Successful immunoprecipitation of dimethyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using ChIP Primers B-Globin.
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 05-1249, Anti-dimethyl H3 (Lys9), clone CMA307 at 2.0ug/mL (1:500) dilution. Proteins were visualized using a Donkey anti-mouse IgG conjugated to HRP and a chemiluminescence detection system.

ELISA:
This antibody has been shown by an outside laboratory to be suitable for ELISA. Kimura, H., et al. (2008). Cell Struct. Funct. 33: 61-73.

Immunocytochemistry:
This antibody has been shown by an outside laboratory to be suitable for immunocytochemistry ( Kimura, 2008).

Immunoprecipitation:
This antibody has been shown by an outside laboratory to be suitable for immunoprecipitation ( Kimura, 2008).

Multiplexing:
This antibody specifically recognizes histone H3 dimethylated on Lys9 by Luminex assay.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Histones
Use Anti-Dimethyl Histone H3 (Lys9) Antibody, clone CMA307 (mouse monoclonal antibody) validated in WB, ELISA, ICC, IP, Mplex to detect Dimethyl Histone H3 (Lys9) also known as H3K9me2, Histone H3 (di methyl K9).

Quality

Western Blot Analysis: 1:10,000 of this lot detected Dimethyl Histone H3 in HeLa acid extract.

Target description

~17 kDa

Linkage

Replaces: 04-791

Physical form

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

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

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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John R Horton et al.
The Journal of biological chemistry, 299(2), 102862-102862 (2023-01-04)
The N-terminal half of PHF2 harbors both a plant homeodomain (PHD) and a Jumonji domain. The PHD recognizes both histone H3 trimethylated at lysine 4 and methylated nonhistone proteins including vaccinia-related kinase 1 (VRK1). The Jumonji domain erases the repressive
Nicole I Bieberstein et al.
Scientific reports, 6, 29961-29961 (2016-07-22)
In search for the function of local chromatin environment on pre-mRNA processing we established a new tool, which allows for the modification of chromatin using a targeted approach. Using Transcription Activator-Like Effector domains fused to histone modifying enzymes (TALE-HME), we
Differential H3K4 methylation identifies developmentally poised hematopoietic genes.
Orford, Keith, et al.
Developmental Cell, 14, 798-809 (2008)
Hui Li et al.
PLoS genetics, 7(6), e1002122-e1002122 (2011-06-23)
Eukaryotic nuclei contain regions of differentially staining chromatin (heterochromatin), which remain condensed throughout the cell cycle and are largely transcriptionally silent. RNAi knockdown of the highly conserved heterochromatin protein HP1 in Drosophila was previously shown to preferentially reduce male viability.
Transcriptional activation of transposable elements in mouse zygotes is independent of Tet3-mediated 5-methylcytosine oxidation.
Inoue, A; Matoba, S; Zhang, Y
Cell research null

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