Diagenode

>>>   Click for Diagenode’s approach to COVID-19

H3K9me3 Antibody - ChIP Grade (sample size)

Catalog Number
Format
Price
C15200153-10
(MAb-153-050)
10 µg
$95.00
  Bulk order
Other format



Monoclonal antibody raised in mouse against histone H3, trimethylated at lysine 9 (H3K9me3), using a KLH-conjugated synthetic peptide. 

Lot001
Concentration1.0 µg/µl
Species reactivityHuman, fungi
TypeMonoclonal
PurityProtein A purified
Host
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 1-3 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

    H3K9me3 Antibody ChIP Grade

    H3K9me3 Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K9me3
    A. ChIP was performed using HeLa cells, the monoclonal antibody against H3K9me3 (Cat. No. MAb-153-050) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode Bioruptor using the “Shearing ChIP” kit (Cat. No. kch- redmod-100). ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), 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.
    B. ChIP was performed with the “iDeal ChIP” kit (Cat. No. AB-001-0024), using sheared chromatin from 1 million K562 cells. A titration of the antibody consisting of 1, 2 and 5 μg per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the GAPDH and EIF4A2 genes, used as negative controls, and for the ZNF12 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    H3K9me3 Antibody ELISA validation

    Figure 2. Cross reactivity of the Diagenode monoclonal antibody directed against H3K9me3
    To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K9me3 (Cat. No. MAb-153-050). The wells were coated with peptides containing the unmodified H3K9 as well as the mono-, di- and trimethylated H3K9 and the trimethylated H3K4 and H3K27. Figure 2 shows a high specificity of the antibody for the modification of interest.

    H3K9me3 Antibody validated in Western Blot

    Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H3K9me3
    Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K9me3 (Cat. No. MAb-153-050) 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.

    H3K9me3 Antibody validated in Immunofluorescence

    Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K9me3
    HeLa cells were stained with the Diagenode antibody against H3K9me3 (Cat. No. MAb-153-050) 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 H3K9me3 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.

  • Target Description

    Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases.

  •  Applications
    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
    ELISA
    Enzyme-linked immunosorbent assay. 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
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
    Download
    Datasheet H3K9me3 C15200153 DATASHEET
    Datasheet description
    Download
  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K9me3 Antibody - ChIP Grade (sample size) (Diagenode Cat# C15200153-10 Lot# 001). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Membrane-Bound Methyltransferase Complex VapA-VipC-VapB Guides Epigenetic Control of Fungal Development.
    Sarikaya-Bayram O, Bayram O, Feussner K, Kim JH, Kim HS, Kaever A, Feussner I, Chae KS, Han DM, Han KH, Braus GH
    Epigenetic and transcriptional control of gene expression must be coordinated in response to external signals to promote alternative multicellular developmental programs. The membrane-associated trimeric complex VapA-VipC-VapB controls a signal transduction pathway for fungal differentiation. The VipC-VapB methyltra...

       Site map   |   Contact us   |   Conditions of sales   |   Conditions of purchase   |   Privacy policy   |   Diagenode Diagnostics