Diagenode

H3K9me1 polyclonal antibody - Classic

RFX5-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15410065
(pAb-065-050)
50 µg/43 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.

LotA89-0041
Concentration1.17 µg/µl
Species reactivityHuman, Nematodes
TypePolyclonal
PurityAffinity purified
HostRabbit
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 1 μg/ChIP Fig 1
ELISA 1:500 - 1:1,000 Fig 2
Dot Blotting 1:20,000 Fig 3
Western Blotting 1:1,000 Fig 4
* Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.
  • Validation Data

    ChIP results

    Figure 1 ChIP results obtained with the Diagenode antibody directed against H3K9me1
    ChIP assays were performed using human HeLa cells, the Diagenode antibody directed against H3K9me1 (Cat. No. pAb-065-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016), using sheared chromatin from 10,000 cells. Respectively 1 and 5 μg of the antibody and 5 μg of IgG (negative IP control) were used per ChIP experiment. QPCR was performed with primers for the GAPDH promoter and for the inactive gene MYOD. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K9me1 is preferably present at silent genes.

    ELISA

    Figure 2 Determination of the antibody titer
    To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against human H3K9me1 (Cat. No. pAb-065-050), crude serum and flow through in antigen coated wells. 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 1:68,000.

    Dot Blot

    Figure 3 Cross reactivity test of 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. pAb-065-050) with peptides containing other modifications and the unmodified sequence of histone H3. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. 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 (15 μg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K9me1 (Cat. No. pAb-065-050) 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.

  • 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
    ChIP-qPCR (ab)
    Read more
  • Documents
    Datasheet H3K9me1 pAb-065-050 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
  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K9me1 polyclonal antibody - Classic (Diagenode Cat# C15410065 Lot# A89-0041). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    The histone demethylase JMJD2A/KDM4A links ribosomal RNA transcription to nutrients and growth factors availability
    Salifou K, Ray S, Verrier L, Aguirrebengoa M, Trouche D, Panov KI, Vandromme M
    The interplay between methylation and demethylation of histone lysine residues is an essential component of gene expression regulation and there is considerable interest in elucidating the roles of proteins involved. Here we report that histone demethylase KDM4A/JMJD2A, which is involved in the regulation of cell pr...

    Germline organization in Strongyloides nematodes reveals alternative differentiation and regulation mechanisms.
    Kulkarni A et al.
    Nematodes of the genus Strongyloides are important parasites of vertebrates including man. Currently, little is known about their germline organization or reproductive biology and how this influences their parasitic life strategies. Here, we analyze the structure of the germline in several Strongyloides and closely ...

    The histone demethylase enzyme KDM3A is a key estrogen receptor regulator in breast cancer.
    Wade MA, Jones D, Wilson L, Stockley J, Coffey K, Robson CN, Gaughan L
    Endocrine therapy has successfully been used to treat estrogen receptor (ER)-positive breast cancer, but this invariably fails with cancers becoming refractory to treatment. Emerging evidence has suggested that fluctuations in ER co-regulatory protein expression may facilitate resistance to therapy and be involved i...

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