H3K9me2 Antibody
Monoclonal antibody raised in mouse against histone H3, dimethylated at lysine 9 (H3K9me2), using a KLH-conjugated synthetic peptide.
| Lot | 001-013 |
|---|---|
| Concentration | 1.0 µg/µl |
| Species reactivity | Human, zebrafish, Arabidopsis, silena latifolia: positive. Other species: not tested. |
| Type | Monoclonal |
| Purity | Protein A purified monoclonal antibody. |
| Host | Mouse |
| Storage Conditions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | PBS containing 0.05% azide and 0.05% ProClin 300. |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| ChIP * | 3 µg/ChIP | Fig 1 |
| ELISA | 1:1,000 | Fig 2 |
| Western Blotting | 1:1,000 | Fig 3 |
| Immunofluorescence | 1:500 | Fig 4 |
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 µg per IP.
Validation Data
Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K9me2
ChIP assays were performed using HeLa cells, the monoclonal antibody against H3K9me2 (Cat. No. C15200154) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode Bioruptor using the “Shearing ChIP” kit (Cat. No. C01020021). ChIP was performed with the “OneDay ChIP” kit (Cat. No. C01010080), using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 1, 3 and 9 µg per ChIP experiment was analysed. IgG (5 µg/IP) was used as negative IP control. QPCR was performed with primers for the promoter and the coding region of the GAPDH gene, and for the RPL10 and HBB promoters. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Figure 2. Cross reactivity of the Diagenode monoclonal antibody directed against H3K9me2
To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K9me2 (cat. No. C15200154). The wells were coated with peptides containing the unmodified H3K9 as well as the mono-, di- and trimethylated H3K9 and the dimethylated H3K27. Figure 2 shows a high specificity of the antibody for the modification of interest.
Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H3K9me2
Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K9me2 (Cat. No. C15200154) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K9me2
HeLa cells were stained with the Diagenode antibody against H3K9me2 (Cat. No. C15200154) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9me2 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
Applications
| ELISA Enzyme-linked immunosorbent assay. Read more |
| WB Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more |
| IF Immunofluorescence: Diagenode offers huge selection of highly sensitive antibodies validated in IF. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 HeLa cells transfected with a Cas9 expression vector (... Read more |
| ChIP-qPCR (ab) Read more |
Documents
| Datasheet H3K9me2 C15200154 DATASHEET Datasheet description | Download |
| Antibodies you can trust POSTER Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar... | Download |
| Epigenetic Antibodies Brochure BROCHURE More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e... | Download |
Safety sheets
Publications
How to properly cite our product/service in your workWe strongly recommend using this: H3K9me2 Antibody (Hologic Diagenode Cat# C15200154 Lot# 001-013). Click here to copy to clipboard. Using our products or services in your publication? Let us know! |
Histone lysine demethylase inhibition reprograms prostate cancermetabolism and mechanics. |
Histone post-translational modifications in Silene latifolia X and Y chromosomes suggest a mammal-like dosage compensation system |
The histone methyltransferase DOT1L is required for proper DNA damage response, DNA repair, and modulates chemotherapy responsiveness. |
Role of Annexin gene and its regulation during zebrafish caudal fin regeneration |
Independent Mechanisms Target SMCHD1 to Trimethylated Histone H3 Lysine 9-Modified Chromatin and the Inactive X Chromosome |
A gene expression signature identifying transient DNMT1 depletion as a causal factor of cancer-germline gene activation in melanoma |
Epigenetic hierarchy within the MAGEA1 cancer-germline gene: promoter DNA methylation dictates local histone modifications. |