Diagenode

H4K20me3 polyclonal antibody - Classic

RFX5-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15310057
(CS-057-100)
100 µl
$295.00
  Bulk order
OUT OF STOCK

As an alternative we offer the purified H4K20me3 polyclonal antibody (C15410057).

Polyclonal antibody raised in rabbit against histone H4 containing the trimethylated lysine 20 (H4K20me3), using a KLH-conjugated synthetic peptide.

LotA9-002
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 1:100 Fig 1
ELISA 1:50 Fig 2
Dot Blotting 1:1,000 Fig 3
Western Blotting 1:750 Fig 4
  • Validation data

    chip

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K20me3
    ChIP assays were performed using undifferentiated human teratocarcinoma cells (NCCIT), the Diagenode antibody against H4K20me3 (cat. No. CS-057-100) and optimized PCR primer sets for qPCR. Sheared chromatin from 10,000 cells was used per ChIP experiment. The antibody was tested at two different dilutions of 1:500 and 1:100. The pre-immune serum (PI) was used as a negative control. Quantitative PCR was performed using primer sets for the satellite repeat Sat2 as a positive control and for the promoter of the house keeping gene c-fos, as a negative control. 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 H4K20me3 is preferably present at heterochromatin.

    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 H4K20me3 (cat. No. CS-057-100). 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 1:700.

    dot blot

    Figure 3. Cross reactivity test using the Diagenode antibody directed against H4K20me3
    Dot Blot was used to check the specificity of the Diagenode antibody against H4K20me3 with peptides containing other modifications of histone H3 and H4. Other histone modifications include monoand dimethylation of the same lysine and mono-, di- and trimethylation of lysines 9, 27 and 36 of H3. 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 1:1,000. 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 H4K20me3
    Histone (acid) extracts of NB4 cells were analysed by Western blot using the Diagenode antibody against H4K20me3, diluted 1:750 in TBS-Tween containing 5% skimmed milk. The location of the protein of interest is indicated on the right.

  • 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 H4K20me3 CS-057-100 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...
    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: H4K20me3 polyclonal antibody - Classic (Diagenode Cat# C15310057 Lot# A9-002 ). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Histone modifications at the blastocyst Axin1(Fu) locus mark the heritability of in vitro culture-induced epigenetic alterations in mice.
    Fernandez-Gonzalez R, Ramirez MA, Pericuesta E, Calle A, Gutierrez-Adan A
    For epigenetic phenotypes to be passed on from one generation to the next, it is required that epigenetic marks between generations are not cleared during the two stages of epigenetic reprogramming: mammalian gametogenesis and preimplantation development. The molecular nature of the chromatin marks involved in these...

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