Diagenode

EZH2 polyclonal antibody - Classic

Histone-Deacetylase-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15410039
(pAb-039-050)
50 µg/50 µl
$295.00
  Bulk order

Alternative names: ENX-1, KMT6, EZH1

Polyclonal antibody raised in rabbit against the N-terminus (aa1-343) of the mouse EZH2 protein (Enhancer of zeste homolog 2).

Lot003
Concentration1.0 µg/µl
Species reactivityHuman, mouse
TypePolyclonal
PurityProtein G purified
HostRabbit
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 2 μg/ChIP Fig 1, 2
Western Blotting 1:1,000 Fig 3
Immunochemistry 1:100 - 1:500 Ref 1

* Please note that of 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 EZH2
    ChIP assays were performed using K562 cells, the Diagenode antibody against EZH2 (Cat. No. C15410039) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2. 5 and 10 μg per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. Quantitative PCR was performed with primers for MYT1 and HOXA9, used as positive control targets, and for the coding regions of the active CCT5 and EIF2S3 genes, 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).

    ChIP-seq figure A

    ChIP-seq figure B

    ChIP-seq figure C

    ChIP-seq figure D

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against EZH2
    ChIP was performed on sheared chromatin from 4 million K562 cells using 2 μg of the Diagenode antibody against EZH2 (Cat. No. C15410039) as described above. The IP’d DNA was subsequently 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 short arm and a 6 Mb region containing several enriched regions of human chromosome 3 (figure 2A and B, respectively), and in two genomic regions containing the MYT1 gene on chromosome 20 and the HOX cluster on chromosome 7 (figure 2C and D).

    Western blot

    Figure 3. Western blot analysis using the Diagenode antibody directed against EZH2
    Nuclear extracts of HeLa cells (40 μg) were analysed by Western blot using the Diagenode antibody against EZH2 (Cat. No. pAb-039-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest (expected size 85 kDa) is indicated on the right; the marker (in kDa) is shown on the left.

  • Applications
    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
    IC
    Immunochemistry Read more
    ChIP-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
  • Documents
    Datasheet EZH2 pAb-039-050 DATASHEET
    Polyclonal antibody raised in rabbit against the N-terminus (aa1-343) of the mouse EZH2 protein (...
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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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    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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  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: EZH2 polyclonal antibody - Classic (Diagenode Cat# C15410039 Lot# 003). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    H3K4 acetylation, H3K9 acetylation and H3K27 methylation in breast tumor molecular subtypes
    Judes G et al.
    AIM: Here, we investigated how the St Gallen breast molecular subtypes displayed distinct histone H3 profiles. PATIENTS & METHODS: 192 breast tumors divided into five St Gallen molecular subtypes (luminal A, luminal B HER2-, luminal B HER2+, HER2+ and basal-like) were evaluated for their histone H3 modifica...

    Premalignant SOX2 overexpression in the fallopian tubes of ovarian cancer patients: Discovery and validation studies
    Hellner K et al.
    Current screening methods for ovarian cancer can only detect advanced disease. Earlier detection has proved difficult because the molecular precursors involved in the natural history of the disease are unknown. To identify early driver mutations in ovarian cancer cells, we used dense whole genome sequencing of micro...

    MicroRNAs of the miR-290-295 Family Maintain Bivalency in Mouse Embryonic Stem Cells
    Graham B et al.
    Numerous developmentally regulated genes in mouse embryonic stem cells (ESCs) are marked by both active (H3K4me3)- and polycomb group (PcG)-mediated repressive (H3K27me3) histone modifications. This bivalent state is thought to be important for transcriptional poising, but the mechanisms that regulate bivalent genes...

    BRCA1 positively regulates FOXO3 expression by restricting FOXO3 gene methylation and epigenetic silencing through targeting EZH2 in breast cancer
    C Gong, S Yao, A R Gomes, E P S Man, H J Lee, G Gong, S Chang, S-B Kim, K Fujino, S-W Kim, S K Park, J W Lee, M H Lee, KOHBRA study group, U S Khoo and E W-F Lam
    BRCA1 mutation or depletion correlates with basal-like phenotype and poor prognosis in breast cancer but the underlying reason remains elusive. RNA and protein analysis of a panel of breast cancer cell lines revealed that BRCA1 deficiency is associated with downregulation of the expression of the pleiotropic tumour ...

    The JMJD3 Histone Demethylase and the EZH2 Histone Methyltransferase in Prostate Cancer
    Daures M, Ngollo M, Judes G, Rifaï K, Kemeny JL, Penault-Llorca F, Bignon YJ, Guy L, Bernard-Gallon D
    Prostate cancer is themost common cancer in men. It has been clearly established that genetic and epigenetic alterations of histone 3 lysine 27 trimethylation (H3K27me3) are common events in prostate cancer. This mark is deregulated in prostate cancer (Ngollo et al., 2014). Furthermore, H3K27me3 levels are determine...

    Spatial Interplay between Polycomb and Trithorax Complexes Controls Transcriptional Activity in T Lymphocytes
    Onodera A, Tumes DJ, Watanabe Y, Hirahara K, Kaneda A, Sugiyama F, Suzuki Y, Nakayama T
    Trithorax group (TrxG) and Polycomb group (PcG) proteins are two mutually antagonistic chromatin modifying complexes, however, how they together mediate transcriptional counter-regulation remains unknown. Genome-wide analysis revealed that binding of Ezh2 and menin, central members of the PcG and TrxG complexes, res...

    Loss of EZH2 results in precocious mammary gland development and activation of STAT5-dependent genes
    Yoo KH, Oh S, Kang K, Hensel T, Robinson GW, Hennighausen L
    Establishment and differentiation of mammary alveoli during pregnancy are controlled by prolactin through the transcription factors STAT5A and STAT5B (STAT5), which also regulate temporal activation of mammary signature genes. This study addressed the question whether the methyltransferase and transcriptional co-act...

    Polycomb repressive complex PRC1 spatially constrains the mouse embryonic stem cell genome
    Schoenfelder S et al.
    The Polycomb repressive complexes PRC1 and PRC2 maintain embryonic stem cell (ESC) pluripotency by silencing lineage-specifying developmental regulator genes. Emerging evidence suggests that Polycomb complexes act through controlling spatial genome organization. We show that PRC1 functions as a master regulator of m...

    H19 lncRNA controls gene expression of the Imprinted Gene Network by recruiting MBD1.
    Monnier P, Martinet C, Pontis J, Stancheva I, Ait-Si-Ali S, Dandolo L
    The H19 gene controls the expression of several genes within the Imprinted Gene Network (IGN), involved in growth control of the embryo. However, the underlying mechanisms of this control remain elusive. Here, we identified the methyl-CpG-binding domain protein 1 MBD1 as a physical and functional partner of the H19 ...

    A key role for EZH2 in epigenetic silencing of HOX genes in mantle cell lymphoma.
    Kanduri M, Sander B, Ntoufa S, Papakonstantinou N, Sutton LA, Stamatopoulos K, Kanduri C, Rosenquist R
    The chromatin modifier EZH2 is overexpressed and associated with inferior outcome in mantle cell lymphoma (MCL). Recently, we demonstrated preferential DNA methylation of HOX genes in MCL compared with chronic lymphocytic leukemia (CLL), despite these genes not being expressed in either entity. Since EZH2 has been s...

    The polycomb protein Ezh2 regulates differentiation and plasticity of CD4(+) T helper type 1 and type 2 cells.
    Tumes DJ, Onodera A, Suzuki A, Shinoda K, Endo Y, Iwamura C, Hosokawa H, Koseki H, Tokoyoda K, Suzuki Y, Motohashi S, Nakayama T
    After antigen encounter by CD4(+) T cells, polarizing cytokines induce the expression of master regulators that control differentiation. Inactivation of the histone methyltransferase Ezh2 was found to specifically enhance T helper 1 (Th1) and Th2 cell differentiation and plasticity. Ezh2 directly bound and facilitat...

    Histone lysine trimethylation or acetylation can be modulated by phytoestrogen, estrogen or anti-HDAC in breast cancer cell lines.
    Dagdemir A, Durif J, Ngollo M, Bignon YJ, Bernard-Gallon D
    AIM: The isoflavones genistein, daidzein and equol (daidzein metabolite) have been reported to interact with epigenetic modifications, specifically hypermethylation of tumor suppressor genes. The objective of this study was to analyze and understand the mechanisms by which phytoestrogens act on chromatin in breast c...

    Dynamic Changes in Ezh2 Gene Occupancy Underlie Its Involvement in Neural Stem Cell Self-Renewal and Differentiation towards Oligodendrocytes
    Sher F, Boddeke E, Olah M, Copray S
    Background: The polycomb group protein Ezh2 is an epigenetic repressor of transcription originally found to prevent untimely differentiation of pluripotent embryonic stem cells. We previously demonstrated that Ezh2 is also expressed in multipotent neural stem cells (NSCs). We showed that Ezh2 expression is downregul...

    RYBP-PRC1 Complexes Mediate H2A Ubiquitylation at Polycomb Target Sites Independently of PRC2 and H3K27me3.
    Tavares L, Dimitrova E, Oxley D, Webster J, Poot R, Demmers J, Bezstarosti K, Taylor S, Ura H, Koide H, Wutz A, Vidal M, Elderkin S, Brockdorff N
    Polycomb-repressive complex 1 (PRC1) has a central role in the regulation of heritable gene silencing during differentiation and development. PRC1 recruitment is generally attributed to interaction of the chromodomain of the core protein Polycomb with trimethyl histone H3K27 (H3K27me3), catalyzed by a second complex...

    PcG complexes set the stage for epigenetic inheritance of gene silencing in early S phase before replication.
    Lanzuolo C, Lo Sardo F, Diamantini A, Orlando V
    Polycomb group (PcG) proteins are part of a conserved cell memory system that conveys epigenetic inheritance of silenced transcriptional states through cell division. Despite the considerable amount of information about PcG mechanisms controlling gene silencing, how PcG proteins maintain repressive chromatin during ...

    Silencing of Kruppel-like factor 2 by the histone methyltransferase EZH2 in human cancer.
    Taniguchi H, Jacinto FV, Villanueva A, Fernandez AF, Yamamoto H, Carmona FJ, Puertas S, Marquez VE, Shinomura Y, Imai K, Esteller M
    The Kruppel-like factor (KLF) proteins are multitasked transcriptional regulators with an expanding tumor suppressor function. KLF2 is one of the prominent members of the family because of its diminished expression in malignancies and its growth-inhibitory, pro-apoptotic and anti-angiogenic roles. In this study, we ...

    Enhancer of Zeste 2 (EZH2) is up-regulated in malignant gliomas and in glioma stem-like cells.
    Orzan F, Pellegatta S, Poliani PL, Pisati F, Caldera V, Menghi F, Kapetis D, Marras C, Schiffer D, Finocchiaro G
    AIMS: Proteins of the Polycomb repressive complex 2 (PRC2) are epigenetic gene silencers and are involved in tumour development. Their oncogenic function might be associated with their role in stem cell maintenance. The histone methyltransferase Enhancer of Zeste 2 (EZH2) is a key member of PRC2 function: we have in...

    Promoter-exon relationship of H3 lysine 9, 27, 36 and 79 methylation on pluripotency-associated genes.
    Barrand S, Andersen IS, Collas P
    Evidence links pluripotency to a gene regulatory network organized by the transcription factors Oct4, Nanog and Sox2. Expression of these genes is controlled by epigenetic modifications on regulatory regions. However, little is known on profiles of trimethylated H3 lysine residues on coding regions of these genes in...

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