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H3K4me1 Antibody - ChIP-seq Grade

Catalog Number
Format
Price
C15210018
100 µg
$400.00
  Bulk order



Monoclonal antibody raised in rabbit against the region of histone H3 containing the monomethylated lysine 4 (H3K4me1), using a KLH-conjugated synthetic peptide.

Lot001
Concentration1 μg/μl
Species reactivityHuman, wide range expected.
TypeMonoclonal ChIP grade, ChIP-seq grade
PurityAffinity purified polyclonal antibody.
HostRabbit
Storage ConditionsStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
Storage BufferPBS containing 50% glycerol, 1% BSA and 0.09% azide.
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP/ChIP-seq * 0.5 - 1 µg per IP Fig 1, 2
Dot Blotting 1:2,000 Fig 3
Western Blotting 1:1,000 Fig 4
Immunofluorescence 1:500 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

    Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me1
    ChIP was performed with the Diagenode antibody against H3K4me1 (Cat. No. C15210018) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for a region surrounding the ACTB and GAS2L1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    A.

    B.

    C.

    D.

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K4me1
    ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 0.5 µg of the Diagenode antibody against H3K4me1 (Cat. No. C15210018) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 H3K4me1 signal along the complete sequence and 1.5 Mb region of human chromosome 1 (figure 2A and B), and in two genomic regions containing the ACTB and GAS2L1 positive controls (figure 2C and D).

    Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K4me1
    To test the cross reactivity of the Diagenode antibody against H3K4me1 (Cat. No. C15210018), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 3 shows a high specificity of the antibody for the modification of interest.

    Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me1
    Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H3K4me1 (Cat. No. C15210018). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.

    Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me1
    HeLa cells were stained with the Diagenode antibody against H3K4me1 (Cat. No. C15210018, red) diluted 1:1,000. Actin was stained with fluorescein phalladoin (green).

  •  Applications
    ChIP-qPCR (ab)
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    ChIP-seq (ab)
    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
    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
  •  Documents
    Datasheet H3K4me1 C15210018 DATASHEET
    Monoclonal antibody raised in rabbit against the region of histone H3 containing the monomethylat...
    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

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