Diagenode

H3K4me3 polyclonal antibody - Classic

RFX5-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15410203
(CSP-003-050)
50 µl
$295.00
  Bulk order

Polyclonal antibody raised in rabbit against the region of histone H3 containing the trimethylated lysine 4 (H3K4me3), using a KLH-conjugated synthetic peptide.

LotA.57-0032
Concentrationnot determined
Species reactivityHuman
TypePolyclonal
PurityAffinity purified
HostRabbit
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 3 μl/IP Fig 1
ELISA 1:50 Fig 2
Dot Blotting 1:10,000 Fig 3
Western Blotting 1:250 Fig 4

* Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.1-5 μl per IP.

  • Validation Data

    ChIP

    Figure 1 ChIP results obtained with the Diagenode antibody directed against H3K4me3
    ChIP assays were performed using human U2OS cells, the Diagenode antibody against H3K4me3 (Cat. No. CSP-003-050) and optimized PCR primer pairs for qPCR. ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), using sheared chromatin from 2 million cells and stringent washing conditions. A titration consisting of 0.1, 1 and 3 μl of antibody per ChIP experiment was analyzed. IgG (1 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the promoter of the constitutively expressed GAPDH gene and for myoglobin exon 2. 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 trimethylation of K4 at histone H3 is associated with the promoters of active genes.

    ELISA

    Figure 2 Determination of the antibody titer
    To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody against H3K4me3 (Cat. No. CSP-003-050), the crude serum and the supernatant in antigen coated wells. 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 1:5,250.

    Dot Blot

    Figure 3 Cross reactivity tests using the Diagenode antibody directed against H3K4me3
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4me3 (Cat. No. CSP-003-050) with peptides containing other H3K4 methylations and the unmodified sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:10,000. 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 H3K4me3
    Histone extracts from HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K4me3 (Cat. No. CSP-003-050) diluted 1:250 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 H3K4me3 CSP-003-100 DATASHEET
    Datasheet description
    Download
  • Publications

    How to properly cite this product in your work

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