Diagenode

H4K20me1 monoclonal antibody - Classic (sample size)

Histone-Deacetylase-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15200147-10
(MAb-147-100)
10 µg
$80.00
  Bulk order
Other format

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

Lot003
Concentration2.8 μg/μl
Species reactivityHuman, mouse
TypeMonoclonal
PurityProtein A purified
HostMouse
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 0,5 - 1 μg/ChIP Fig 1, 2
Dot Blotting 1:20,000 Fig 3
Western Blotting 1:1,000 Fig 4
Immunofluorescence 1:200 Fig 5
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5-5 μg per IP.
  • Validation Data

    ChIP

    Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1
    ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    ChIP-seq

    ChIP-seq

    ChIP-seq

    ChIP-seq

    Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1
    ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively).

    Dot Blot

    Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1
    To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. 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 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.

    Western Blot

    Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1
    Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right

    Immunofluorescence

    Immunofluorescence

    Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1
    Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl).

  • Applications
    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-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
  • Documents
    Datasheet H4K20me1 C15200147 DATASHEET
    Datasheet description
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    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...
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  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H4K20me1 monoclonal antibody - Classic (sample size) (Diagenode Cat# C15200147-10 Lot# 003). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    SET8 methyltransferase activity during the DNA double-strand break response is required for recruitment of 53BP1
    Dulev, S., Tkach, J., Lin, S. and Batada, N. N.
    DNA double-strand breaks (DSBs) activate a signaling pathway known as the DNA damage response (DDR) which via protein–protein interactions and post-translational modifications recruit signaling proteins, such as 53BP1, to chromatin flanking the lesion. Depletion of the SET8 methyltransferase prevents accumulation of...

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