Diagenode

H3K4me3 polyclonal antibody - Classic

Histone-Deacetylase-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15410030
(pAb-030-050)
50 µg/25 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.

Lot001
Concentration2.0 µg/µl
Species reactivityHuman, mouse, Arabidopsis
TypePolyclonal
PurityAffinity purified
HostRabbit
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 1 μg/ChIP Fig 1, 2
Dot Blotting 1:2,000 Fig 3
Western Blotting 1:500 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 1-5 μg per IP.
  • Validation data

    ChIP

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4me3
    ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4me3 (Cat. No. pAb-030-050) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. AB-001-0024), using sheared chromatin from 1 million cells. 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 active genes GAPDH and EIF4A2, 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

    ChIP-seq

    ChIP-seq

    ChIP-seq

    ChIP-seq

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K4me3
    ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 μg of the Diagenode antibody against H3K4me3 (Cat. No. pAb-030-050) 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 the complete sequence and a 600 kb region of the X-chromosome (figure 2A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes, respectively (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.

    Dot Blot

    Figure 3. Cross reactivity test 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. pAb-030-050) with peptides containing other modifications and unmodified sequences of histone H3 and H4. 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.

    Western Blot

    Figure 4. Western blot analysis using the Diagenode antibody directed against H3K4me3
    Western blot was performed using histone extracts from HeLa cells (HeLa HE, 15 μg) and the Diagenode antibody against H3K4me3 (Cat. No. pAb-030-050) diluted 1:500 in TBS-Tween containing 5% skimmed milk. A molecular weight marker (in kDa) is shown on the left, the position of the protein of interest is shown on the right.

    Immunofluorescence

    Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K4me3
    HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. pAb-030-050) 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 H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

  • Applications
    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-qPCR (ab)
    Read more
    ChIP-seq (ab)
    Read more
  • Documents
    Datasheet H3K4me3 C15410030 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
  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3K4me3 polyclonal antibody - Classic (Diagenode Cat# C15410030 Lot# 001). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Deep sequencing and de novo assembly of the mouse oocyte transcriptome define the contribution of transcription to the DNA methylation landscape
    Veselovska L, Smallwood SA, Saadeh H, Stewart KR, Krueger F, Maupetit-Méhouas S, Arnaud P, Tomizawa S, Andrews S, Kelsey G
    BACKGROUND: Previously, a role was demonstrated for transcription in the acquisition of DNA methylation at imprinted control regions in oocytes. Definition of the oocyte DNA methylome by whole genome approaches revealed that the majority of methylated CpG islands are intragenic and gene bodies are hypermethylated...

    CpG signalling, H2A.Z/H3 acetylation and microRNA-mediated deferred self-attenuation orchestrate foetal NOS3 expression.
    Postberg J, Kanders M, Forcob S, Willems R, Orth V, Hensel KO, Weil PP, Wirth S, Jenke AC
    BACKGROUND: An adverse intrauterine environment leads to permanent physiological changes including vascular tone regulation, potentially influencing the risk for adult vascular diseases. We therefore aimed to monitor responsive NOS3 expression in human umbilical artery endothelial cells (HUAEC) and to study the unde...

    Paclitaxel resistance increases oncolytic adenovirus efficacy via upregulated CAR expression and dysfunctional cell cycle control.
    Ingemarsdotter CK, Tookman LA, Browne A, Pirlo K, Cutts R, Chelela C, Khurrum KF, Leung EY, Dowson S, Webber L, Khan I, Ennis D, Syed N, Crook TR, Brenton JD, Lockley M, McNeish IA
    Resistance to paclitaxel chemotherapy frequently develops in ovarian cancer. Oncolytic adenoviruses are a novel therapy for human malignancies that are being evaluated in early phase trials. However, there are no reliable predictive biomarkers for oncolytic adenovirus activity in ovarian cancer. We investigated the ...

    p53-Independent regulation of p21Waf1/Cip1 expression and senescence by PRMT6.
    Phalke S, Mzoughi S, Bezzi M, Jennifer N, Mok WC, Low DH, Thike AA, Kuznetsov VA, Tan PH, Voorhoeve PM, Guccione E
    p21 is a potent cyclin-dependent kinase inhibitor that plays a role in promoting G1 cell cycle arrest and cellular senescence. Consistent with this role, p21 is a downstream target of several tumour suppressors and oncogenes, and it is downregulated in the majority of tumours, including breast cancer. Here, we repor...

    Transcription and histone methylation changes correlate with imprint acquisition in male germ cells.
    Henckel A, Chebli K, Kota SK, Arnaud P, Feil R
    Genomic imprinting in mammals is controlled by DNA methylation imprints that are acquired in the gametes, at essential sequence elements called 'imprinting control regions' (ICRs). What signals paternal imprint acquisition in male germ cells remains unknown. To address this question, we explored histone methylation ...

  • Related products

Events

  • Workshop on Chromatin Proteomics
    Crete, Greece
    Oct 3-Oct 8, 2016
  • 2nd Annual Next Generation Sequencing Congress
    Boston, USA
    Oct 3-Oct 4, 2016
  • XXXIX Reunión anual de la Sociedad de Bioquimica y Biología Molecular de Chile
    Puerto Varas, Chile
    Sep 27-Sep 30, 2016
 See all events

Twitter feed

News

 See all news