Diagenode

H3K4un Antibody

Catalog Number
Format
Price
C15200149
(MAb-149-050)
50 µg/50 µl
$380.00
  Bulk order
Other format



Monoclonal antibody raised in mouse against histone H3 using a KLH-conjugated synthetic peptide containing the unmodified lysine 4 (H3K4un).

Lot001-11
Concentration1.0 µg/µl
Species reactivityHuman
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 * 1-5 μg/ChIP Fig 1
ELISA 1:5,000 Fig 2
Dot Blotting 1:1,000 Fig 3
Immunofluorescence 1:500 Fig 4

* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.

  • Validation Data

    H3K4un Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4un
    ChIP assays were performed using HeLa cells, the monoclonal antibody against H3K4un (Cat. No. MAb-149-050) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode Bioruptor. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016), using sheared chromatin from 105 cells. A titration of the antibody consisting of 1, 5 and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH promoter (Cat. No. pp-1001-050) and for exon 2 of the myoglobin gene (Cat. No. pp-1006-050). Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    H3K4un Antibody ELISA validation

    Figure 2. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4un
    To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4un (Cat. No. MAb-149-050). The wells were coated with peptides containing the unmodified H3K4 region as well as the mono-, di- and trimethylated H3K4. Figure 2 shows a high specificity of the antibody for the unmodified peptide.

    H3K4un Antibody validated in Western Blot

    Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4un
    Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4un (Cat. No. MAb-149-050) 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.

    H3K4un Antibody validated in Immunofluorescence

    Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4un
    HeLa cells were stained with the Diagenode antibody against H3K4un (Cat. No. MAb-149-050) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K4un antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

  • Target Description

    Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases.

  •  Applications
    ELISA
    Enzyme-linked immunosorbent assay. 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 H3K4un MAb-149-050 DATASHEET
    Datasheet description
    Download
  •  Safety sheets
    H3K4un Antibody SDS US en Download
    H3K4un Antibody SDS GB en Download
    H3K4un Antibody SDS BE fr Download
    H3K4un Antibody SDS FR fr Download
    H3K4un Antibody SDS ES es Download
    H3K4un Antibody SDS DE de Download
    H3K4un Antibody SDS JP ja Download
    H3K4un Antibody SDS BE nl Download
  •  Publications

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