Diagenode

H3K4me2 polyclonal antibody (sample size)

Catalog Number
Format
Price
C15410035-10
(pAb-035-050)
10 µg
$115.00
  Bulk order
Other format



Polyclonal antibody raised in rabbit against histone H3 containing the dimethylated lysine 4 (H3K4me2), using a KLH-conjugated synthetic peptide.

LotA936-0023
Concentration1.1 µg/µl
Species reactivityHuman, Arabidopsis: positive. Other species: not tested.
TypePolyclonal
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 0.05% 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/ChIP Fig 1, 2
CUT&Tag 0.5 µg Fig 3
ELISA 1:500 Fig 4
Dot Blotting 1:20,000 Fig 5
Western Blotting 1:1,000 Fig 6

* Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5-5 µg per IP.

  • Validation Data

    H3K4me2 Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4me2
    ChIP was performed with the Diagenode antibody against H3K4me2 (cat. No. C15410035) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). 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. Quantitative PCR was performed with primers for a region upstream of the ACTB and GAPDH promoters, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, 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. H3K4me2 Antibody Cut &

    B. H3K4me2 Antibody for ChIP-seq

    C. H3K4me2 Antibody for ChIP-seq assay

    D. H3K4me2 Antibody validated in ChIP-seq

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K4me2
    ChIP was performed on HeLa cells using 0.5 µg of the Diagenode antibody against H3K4me2 (cat. No. C15410035). The IP'd DNA was 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 along the complete sequence and a 1.5 Mb region of the human X-chromosome (figure 2A and 2B) and in 2 chromosomal regions surrounding the ACTB and GAPDH positive control genes (figure 2C and D, respectively).

    A. H3K4me2 Antibody Cut&Tag

    B. H3K4me2 Antibody Cut&Tag

    Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K4me2
    CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 0.5 µg of the Diagenode antibody against H3K4me2 (cat. No. C15410035) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the EIF2S3 gene on the X-chromosome (figure 3A and B, respectively).

    H3K4me2 Antibody ELISA validation

    Figure 4. Determination of the titer
    To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K4me2 (cat. No. C15410035) 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 4), the titer of the antibody was estimated to be 1:12,000.

    H3K4me2 Antibody validated in Dot Blot

    Figure 5. Cross reactivity test using the Diagenode antibody directed against H3K4me2
    A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4me2 (cat. No. C15410035) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 5 shows a high specificity of the antibody for the modification of interest.

    H3K4me2 Antibody validated in Western Blot

    Figure 6. Western blot analysis using the Diagenode antibody directed against H3K4me2
    Western blot was performed on whole cell extracts (25 µg, lane 1) and on 1 µg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K4me2 (cat. No. C15410035) 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.

  • 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.

  •  実験手法
    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
    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
    CUT&Tag
    CUT&Tagアッセイを成功させるための重要な要素の1つは使用される抗体の品質です。 特異性高い抗体は、目的のタンパク質のみをターゲットとした確実な結果を可能にします。 CUT&Tagで検証済みの抗体のセレクションはこちらからご覧ください。 Read more: Products for CUT&Tag assay Performance of Diagenode's antibodies in CUT&Tag Read more
  •  資料
    Datasheet H3K4me2 pAb-035-050 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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  •  Safety sheets
    H3K4me2 Antibody SDS GB en Download
    H3K4me2 Antibody SDS US en Download
    H3K4me2 Antibody SDS DE de Download
    H3K4me2 Antibody SDS JP ja Download
    H3K4me2 Antibody SDS BE nl Download
    H3K4me2 Antibody SDS BE fr Download
    H3K4me2 Antibody SDS FR fr Download
    H3K4me2 Antibody SDS ES es Download
  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K4me2 polyclonal antibody (sample size) (Diagenode Cat# C15410035-10 Lot# A936-0023). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

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    Simon C. et al.
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    Stielow B. et al.
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    Inhibition of Histone Demethylases LSD1 and UTX Regulates ERα Signaling in Breast Cancer.
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    We identified a regulatory system that acts downstream of Wnt/β-catenin signaling in salivary gland and head and neck carcinomas. We show in a mouse tumor model of K14-Cre-induced Wnt/β-catenin gain-of-function and Bmpr1a loss-of-function mutations that tumor-propagating cells exhibit increased Mll1 activi...

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