Diagenode

H3K9me1 polyclonal antibody - Classic

RFX5-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15310065
(CS-065-100)
100 µl
$295.00
  Bulk order

Polyclonal antibody raised in rabbit against histone H3 containing the monomethylated lysine 9 (H3K9me1), using a KLH-conjugated synthetic peptide.

LotA89-001
Concentrationnot determined
Species reactivityHuman
TypePolyclonal
PurityWhole antiserum
HostRabbit
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 10 μl/ChIP Fig 1
ELISA 1:2,000 - 1:3,000 Fig 2
Dot Blotting 1:200,000 Fig 3
Western Blotting 1:1,000 Fig 4
Immunofluorescence 1:200 Fig 5
* Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 μl per IP.
  • Validation Data

    ChIP

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9me1
    ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K9me1 (cat. No. CS-065-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “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 2, 5, 10 and 15 μl per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. Quantitative PCR was performed using primers for the promoter of the housekeeping gene GAPDH and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    ELISA

    Figure 2. Determination of the titer
    To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K9me1 (cat. No. CS-065-100). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:50,000.

    Dot Blot

    Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9me1
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9me1 (cat. No. CS-065-100) with peptides containing other modifications of histone H3. Other histone modifications include di- and trimethylation of the same lysine and mono-, di- and trimethylation of lysine 27, 36 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:200,000. Figure 3 shows a high specificity of the antibody for the modification of interest.

    Western Blot

    Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9me1
    Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9me1 (cat. No. CS-065-100) diluted 1:1000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.

    IF

    Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9me1
    HeLa cells were stained with the Diagenode antibody against H3K9me1 (cat. No. CS-065-100) and with DAPI. Cells were formaldehyde fixated, permeabilized with Triton X-100 and blocked with PBS containing 2.5% BSA. Figure 5A: cells were immunofluorescently labelled with the H3K9me1 antibody (diluted 1:200 and incubated for 1 hour at room temperature) followed by goat anti-rabbit antibody conjugated to DyLight. Figure 5B: staining of the nuclei with DAPI, which specifically labels DNA. Both antibody and DAPI staining are restricted to the nucleus.

  • Applications
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    DB
    Dot blotting 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 H3K9me1 CS-065-100 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...
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    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
    Download
  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K9me1 polyclonal antibody - Classic (Diagenode Cat# C15310065 Lot# A89-001). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Chromatin states of core pluripotency-associated genes in pluripotent, multipotent and differentiated cells.
    Barrand S, Collas P
    Oct4, Nanog and Sox2 constitute a core of transcription factors controlling pluripotency. Differentiation and reprogramming studies have unraveled a few epigenetic modifications associated in relation to the expression state of OCT4, NANOG and SOX2. There is, however, no comprehensive map of chromatin states on thes...

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