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H3K4me3 Antibody - ChIP-seq Grade

cut and tag antibody icon

目录号
格式
价格
C15200152
(MAb-152-050)
50 µg
$380.00
  Bulk order
其他格式
C15200152-10
10 µg



Monoclonal antibody raised in mouse against histone H3, trimethylated at lysine 4 (H3K4me3), using a KLH-conjugated synthetic peptide.

Lot001-15
Concentration1.0 µg/µl
Species reactivityHuman, mouse, Nematodes, Arabidopsis
TypeMonoclonal
ChIP-grade
ChIP-seq grade
PurityProtein A purified monoclonal antibody.
HostMouse
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 * 2 μg/ChIP Fig 1,2
CUT&TAG 1 μg Fig 3
ELISA 1:3,000 Fig 4
Western Blotting 1:1,000 Fig 5
Immunofluorescence 1:500 Fig 6

* 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

    Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3

    ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.

    A.H3K4me3 Antibody ChIP-seq Grade

    B.H3K4me3 Antibody for ChIP-seq

    C.H3K4me3 Antibody for ChIP-seq assay

    D.H3K4me3 Antibody validated in ChIP-seq

    Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3
    ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.

    A.

    B.

    Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3
    CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) 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 along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).

    Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3
    To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.

    Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3
    Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) 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.

    Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3
    HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 H3K4me3 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.

  •  证明书

    I have extensively used the antibodies against the histone modifications H3K4me3, H3k27me3, H3K9ac, H4k8ac and H3K18ac provided by Diagenode. The high level of specificity and selectivity of these antibodies in mouse brain samples, confirmed by using several negative and positive controls run in parallel with mouse brain tissue samples, ensured successful and reproducible results. I have been a Diagenode costumer for over one year now and I am extremely satisfied with the efficiency of the Bioruptor Pico for chromatin shearing as well as all of the ChIP materials (i.e., antibodies, blocking peptides, primer pairs for qPCR) provided by this company. Many thanks.

    Dr. Ermelinda Lomazzo, Institute of Physiological Chemistry, AG Prof. Beat Lutz. University Medical Center of the Johannes Gutenberg University Mainz, Germany
  •  应用
    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-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
    CUT&Tag
    Read more
  •  文档
    Datasheet H3K4me3 C15200152 DATASHEET
    Datasheet description    		 Download
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...    		 Download
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...    		 Download
  •  Safety sheets
    H3K4me3 Antibody SDS GB en Download
    H3K4me3 Antibody SDS US en Download
    H3K4me3 Antibody SDS DE de Download
    H3K4me3 Antibody SDS JP ja Download
    H3K4me3 Antibody SDS BE nl Download
    H3K4me3 Antibody SDS BE fr Download
    H3K4me3 Antibody SDS FR fr Download
    H3K4me3 Antibody SDS ES es Download
  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K4me3 Antibody - ChIP-seq Grade (Diagenode Cat# C15200152 Lot# 001-15). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Local euchromatin enrichment in lamina-associated domains anticipatestheir repositioning in the adipogenic lineage.
    Madsen-Østerbye J. et al.
    BACKGROUND: Interactions of chromatin with the nuclear lamina via lamina-associated domains (LADs) confer structural stability to the genome. The dynamics of positioning of LADs during differentiation, and how LADs impinge on developmental gene expression, remains, however, elusive. RESULTS: We examined changes in t...

    The glucocorticoid receptor recruits the COMPASS complex to regulateinflammatory transcription at macrophage enhancers.
    Greulich, Franziska et al.
    Glucocorticoids (GCs) are effective anti-inflammatory drugs; yet, their mechanisms of action are poorly understood. GCs bind to the glucocorticoid receptor (GR), a ligand-gated transcription factor controlling gene expression in numerous cell types. Here, we characterize GR's protein interactome and find the SETD1A ...

    Distinct and temporary-restricted epigenetic mechanisms regulate human αβ and γδ T cell development
    Roels J, Kuchmiy A, De Decker M, et al.
    The development of TCRαβ and TCRγδ T cells comprises a step-wise process in which regulatory events control differentiation and lineage outcome. To clarify these mechanisms, we employed RNA-sequencing, ATAC-sequencing and ChIPmentation on well-defined thymocyte subsets that represent the conti...

    High resolution methylation analysis of the HoxA5 regulatory region in different somatic tissues of laboratory mouse during development
    Sinha P. et al.
    Homeobox genes encode a group of DNA binding regulatory proteins whose key function occurs in the spatial-temporal organization of genome during embryonic development and differentiation. The role of these Hox genes during ontogenesis makes it an important model for research. HoxA5 is a member of Hox gene family pla...

    Detection of Protein-Protein Interactions and Posttranslational Modifications Using the Proximity Ligation Assay: Application to the Study of the SUMO Pathway.
    Ristic M. et al.
    The detection of protein-protein interactions by imaging techniques often requires the overexpression of the proteins of interest tagged with fluorescent molecules, which can affect their biological properties and, subsequently, flaw experiment interpretations. The recent development of the proximity ligation assays...

    Methylation of the Sox9 and Oct4 promoters and its correlation with gene expression during testicular development in the laboratory mouse
    Pamnani M et al.
    Sox9 and Oct4 are two important regulatory factors involved in mammalian development. Sox9, a member of the group E Sox transcription factor family, has a crucial role in the development of the genitourinary system, while Oct4, commonly known as octamer binding transcription factor 4, belongs to class V of the trans...

    Germline organization in Strongyloides nematodes reveals alternative differentiation and regulation mechanisms.
    Kulkarni A et al.
    Nematodes of the genus Strongyloides are important parasites of vertebrates including man. Currently, little is known about their germline organization or reproductive biology and how this influences their parasitic life strategies. Here, we analyze the structure of the germline in several Strongyloides and closely ...

    Transcription termination and chimeric RNA formation controlled by Arabidopsis thaliana FPA.
    Duc C, Sherstnev A, Cole C, Barton GJ, Simpson GG
    Alternative cleavage and polyadenylation influence the coding and regulatory potential of mRNAs and where transcription termination occurs. Although widespread, few regulators of this process are known. The Arabidopsis thaliana protein FPA is a rare example of a trans-acting regulator of poly(A) site choice. Analysi...

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