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

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.

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.

Epitope: dimethyl Lys9

Synthetic peptide corresponding to amino acids 1-19 of human Histone H3, dimethylated on Lys9, conjugated to KLH.

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).

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

~17 kDa

Replaces: 04-791

Format: Purified

Mouse monoclonal IgG1κ supernatant in PBS with 0.05% sodium azide.

Protein G Purified

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.

Control
HeLa acid extract

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

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