Diagenode

H4K20me1 polyclonal antibody - Classic

RFX5-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15310034
(CS-034-100)
100 µl
$295.00
  Bulk order

Polyclonal antibody raised in rabbit against histone H4 containing the monomethylated lysine 20 (H4K20me1), using a KLH-conjugated synthetic peptide.

LotA254-001
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 * 5 - 10 μl/ChIP Fig 1
ELISA 1:300 Fig 2
Dot Blotting 1:20,000 Fig 3
Western Blotting 1:750 Fig 4
* 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 H4K20me1
    ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H4K20me1 (cat. No. CS-034-100) 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 with primers for the GAPDH promoter 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 against H4K20me1 (cat. No. CS-034-100). 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 1:8,000.

    Dot Blot

    Figure 3. Cross reactivity test using the Diagenode antibody directed against H4K20me1
    To check the specificity of the Diagenode antibody against H4K20me1 (cat. No. CS-034-050) a Dot Blot was performed with peptides containing other modifications of histone H4 or the unmodified sequence. Other histone modifications include mono- and dimethylation of the same lysine and acetylation of the nearby lysine 16. To determine the cross reactivity, 0.2 to 100 pmol of peptides were spotted on a membrane. Three different peptides for H4K16ac were used. 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 H4K20me1
    Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H4K20me1 (cat. No. CS-034-100) diluted 1:750 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 H4K20me1 CS-034-100 DATASHEET
    Datasheet description
    Download
  • Publications

    How to properly cite this product in your work

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