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17-662 ChIPAb+ EZH2, clone AC22 - ChIP Validated Antibody and Primer Set

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17-662
25 assays  25 assays per set. Recommended use: ~2 μg antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H, M, Vrt, RWB, ChIP
      Description
      Catalogue Number17-662
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ EZH2, clone AC22 - ChIP Validated Antibody and Primer Set
      OverviewAll ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
      The ChIPAb+ EZH2, clone AC22 antibody/primer set includes the EZH2, clone AC22 antibody, a negative control antibody (purified mouse IgG), and qPCR primers which amplify a 135 bp region within the promoter of the human MYT-1 gene. The EZH2, clone AC22 antibody and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of EZH2-associated chromatin.
      Alternate Names
      • Enhancer of zeste homolog 2
      • Lysine N-methyltransferase 6
      • enhancer of zeste 2
      • enhancer of zeste homolog 2
      • ENX-1
      Background InformationEZH2 is a polycomb group (PcG) protein that is a catalytic subunit of the PRC2/EED-EZH2 complex, which methylates 'Lys-9' (H3K9me) and 'Lys-27' (H3K27me) of histone H3, leading to transcriptional repression of the affected target gene. Able to mono-, di- and trimethylate 'Lys-27' of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively. Compared to EZH2-containing complexes, it is more abundant in embryonic stem cells and plays a major role in forming H3K27me3, which is required for embryonic stem cell identity and proper differentiation. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1, CDKN2A and retinoic acid target genes. EZH2 can also methylate non-histone proteins such as the transcription factor GATA4 and the nuclear receptor RORA. Regulates the circadian clock via histone methylation at the promoter of the circadian genes. Essential for the CRY1/2-mediated repression of the transcriptional activation of PER1/2 by the CLOCK-ARNTL/BMAL1 heterodimer; involved in the di and trimethylation of 'Lys-27' of histone H3 on PER1/2 promoters which is necessary for the CRY1/2 proteins to inhibit transcription.
      References
      Product Information
      FormatPurified
      Control
      • Includes negative control mouse IgG antibody and primers specific for of human MYT-1 promoter region.
      PresentationEZH2, clone AC22 (mouse monoclonal IgG2a). One vial containing 50 μg of protein G purified antibody in 50 μL PBS containing 0.05% sodium azide. Store at -20°C.

      Normal Mouse IgG. Two vials, each containing 25 μg purified mouse IgG in 25 μL storage buffer containing 0.1% sodium azide. Store at -20°C.

      ChIP Primers MYT-1 promoter Region. One vial containing 75 μL of 5 μM of each primer specific for the promoter region of human MYT-1. Store at -20°C.
      FOR: ACA AAG GCA GAT ACC CAA CG
      REV: GCA GTT TCA AAA AGC CAT CC
      Quality LevelMQ100
      Applications
      ApplicationEZH2, clone AC22, ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of EZH2.
      Key Applications
      • Western Blotting
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation:
      Representative lot data.
      Sonicated chromatin prepared from HeLa S3 cells (4 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 µg of either a normal mouse IgG or Anti-EZH2, clone AC22 antibody and the Magna ChIP G Kit (Cat. # 17-611).
      Successful immunoprecipitation of EZH2-associated DNA fragments was verified by qPCR using GAPDH promoter (negative) and MYT-1 promoter (positive) primers (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
      Please refer to the EZ-Magna G ChIP™ (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

      Immunoprecipitation Analysis:
      Representative lot data.
      HeLa nuclear cell extract was Immunoprecipitated with either anti-EZH2, clone AC22 or mouse IgG and then resolved by electrophoresis, transferred to PVDF and probed with Cat. # 05-1319 anti-EZH2, clone BD43 (1:1000 dilution).
      Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP (Cat. # AP124P) and a chemiluminescence detection system.
      (Please see figures).
      Lane 1: Hela nuclear extract immunoprecipitate with anti-EZH2, clone AC22.
      Lane 2: Hela nuclear extract immunoprecipitate with IgG.
      Biological Information
      ImmunogenEZH2, clone AC22 purified antibody is made against a GST fusion protein corresponding to amino acids 353-451 of human EZH2.
      Epitopea.a. 353-451
      CloneAC22
      HostMouse
      SpecificityRecognizes EZH2, MW: ~85-100 kDa.
      IsotypeIgG2a
      Species Reactivity
      • Human
      • Mouse
      • Vertebrates
      • Rat
      Species Reactivity NoteHuman, mouse, rat. Not tested in other species. The immunogen sequence is identical in a wide range of animal and plant species, so broad cross-reactivity is expected.
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThis gene encodes a member of the Polycomb-group (PcG) family. PcG family members form multimeric protein complexes, which are involved in maintaining the transcriptional repressive state of genes over successive cell generations. This protein associates with the embryonic ectoderm development protein, the VAV1 oncoprotein, and the X-linked nuclear protein. This protein may play a role in the hematopoietic and central nervous systems. Two transcript variants encoding distinct isoforms have been identified for this gene.
      Gene Symbol
      • EZH2
      • KMT6
      UniProt Number
      UniProt SummaryFUNCTION: Polycomb group (PcG) protein. Catalytic subunit of the PRC2/EED-EZH2 complex, which methylates 'Lys-9' and 'Lys-27' of histone H3, leading to transcriptional repression of the affected target gene. Able to mono-, di- and trimethylate 'Lys-27' of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively. Compared to EZH2-containing complexes, it is more abundant in embryonic stem cells and plays a major role in forming H3K27me3, which is required for embryonic stem cell identity and proper differentiation. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1 and CDKN2A.
      CATALYTIC ACTIVITY: S-adenosyl-L-methionine + histone L-lysine = S-adenosyl-L-homocysteine + histone N(6)-methyl-L-lysine.
      SUBUNIT STRUCTURE: Binds ATRX via the SET domain Probable. Component of the PRC2/EED-EZH2 complex, which includes EED, EZH2, SUZ12, RBBP4 and RBBP7 and possibly AEBP2. The minimum components required for methyltransferase activity of the PRC2/EED-EZH2 complex are EED, EZH2 and SUZ12. The PRC2 complex may also interact with DNMT1, DNMT3A, DNMT3B and PHF1 via the EZH2 subunit and with SIRT1 via the SUZ12 subunit. Interacts with HDAC1 and HDAC2.
      SUBCELLULAR LOCATION: Nucleus.
      TISSUE SPECIFICITY: Expressed in many tissues. Overexpressed in numerous tumor types including carcinomas of the breast, colon, larynx, lymphoma and testis.
      DEVELOPMENTAL STAGE: Expression decreases during senescence of embryonic fibroblasts (HEFs). Expression peaks at the G1/S phase boundary.
      INDUCTION: Expression is induced by E2F1, E2F2 and E2F3. Expression is reduced in cells subject to numerous types of stress including UV-, IR- and bleomycin-induced DNA damage and by activation of TP53/p53.
      PTM: Phosphorylated by AKT1. Phosphorylation by AKT1 reduces methyltransferase activity.
      SEQUENCE SIMILARITIES: Belongs to the histone-lysine methyltransferase family. EZ subfamily.
      Contains 1 SET domain.
      CAUTION: Two variants of the PRC2 complex have been described, termed PRC3 and PRC4. Each of the three complexes may include a different complement of EED isoforms, although the precise sequences of the isoforms in each complex have not been determined. The PRC2 and PRC4 complexes may also methylate 'Lys-26' of histone H1 in addition to 'Lys-27' of histone H3 (Ref.15 and Ref.19), although other studies have demonstrated no methylation of 'Lys-26' of histone H1 by PRC2
      Molecular Weight~85 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceChromatin Immunoprecipitation:
      Sonicated chromatin prepared from HeLa S3 cells (4 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG or Anti-EZH2, clone AC22 antibody and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of EZH2-associated DNA fragments was verified by qPCR using Control Primers (Please see figures).
      Please refer to the EZ-Magna ChIP™ G (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
      Packaging Information
      Material Size25 assays
      Material Package25 assays per set. Recommended use: ~2 μg antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      ChIPAb+ EZH2, clone AC22 - ChIP Validated Antibody and Primer Set SDS

      Title

      Safety Data Sheet (SDS) 

      ChIPAb+ EZH2, clone AC22 - ChIP Validated Antibody and Primer Set Certificates of Analysis

      TitleLot Number
      ChIPAb+ EZH2, clone AC22 2475706
      ChIPAb+ EZH2, clone AC22 - 2366583 2366583
      ChIPAb+ EZH2, clone AC22 - 2397118 2397118
      ChIPAb+ EZH2, clone AC22 - 2436405 2436405
      ChIPAb+ EZH2, clone AC22 - 1971843 1971843
      ChIPAb+ EZH2, clone AC22 - 2037220 2037220
      ChIPAb+ EZH2, clone AC22 - 2196057 2196057
      ChIPAb+ EZH2, clone AC22 - 2246815 2246815
      ChIPAb+ EZH2, clone AC22 - 3157694 3157694
      ChIPAb+ EZH2, clone AC22 - 3304987 3304987

      References

      Reference overviewSpeciesPub Med ID
      Ezh2 mediated H3K27me3 activity facilitates somatic transition during human pluripotent reprogramming.
      Rao, RA; Dhele, N; Cheemadan, S; Ketkar, A; Jayandharan, GR; Palakodeti, D; Rampalli, S
      Scientific reports  5  8229  2015

      Show Abstract
      25648270 25648270
      YY1 DNA binding and interaction with YAF2 is essential for Polycomb recruitment.
      Basu, A; Wilkinson, FH; Colavita, K; Fennelly, C; Atchison, ML
      Nucleic acids research  42  2208-23  2014

      Show Abstract
      Human24285299 24285299
      JARID2 is involved in transforming growth factor-beta-induced epithelial-mesenchymal transition of lung and colon cancer cell lines.
      Tange, S; Oktyabri, D; Terashima, M; Ishimura, A; Suzuki, T
      PloS one  9  e115684  2014

      Show Abstract
      25542019 25542019
      E2f6-mediated repression of the meiotic Stag3 and Smc1β genes during early embryonic development requires Ezh2 and not the de novo methyltransferase Dnmt3b.
      Leseva, M; Santostefano, KE; Rosenbluth, AL; Hamazaki, T; Terada, N
      Epigenetics  8  873-84  2013

      Show Abstract
      23880518 23880518
      BRG1 promotes survival of UV-irradiated melanoma cells by cooperating with MITF to activate the melanoma inhibitor of apoptosis gene.
      Saladi, SV; Wong, PG; Trivedi, AR; Marathe, HG; Keenen, B; Aras, S; Liew, ZQ; Setaluri, V; de la Serna, IL
      Pigment cell & melanoma research  26  377-91  2013

      Show Abstract
      23480510 23480510
      Alcohol-induced epigenetic alterations to developmentally crucial genes regulating neural stemness and differentiation.
      Veazey, KJ; Carnahan, MN; Muller, D; Miranda, RC; Golding, MC
      Alcoholism, clinical and experimental research  37  1111-22  2013

      Show Abstract
      23488822 23488822
      Redistribution of H3K27me3 upon DNA hypomethylation results in de-repression of Polycomb target genes.
      Reddington, JP; Perricone, SM; Nestor, CE; Reichmann, J; Youngson, NA; Suzuki, M; Reinhardt, D; Dunican, DS; Prendergast, JG; Mjoseng, H; Ramsahoye, BH; Whitelaw, E; Greally, JM; Adams, IR; Bickmore, WA; Meehan, RR
      Genome biology  14  R25  2013

      Show Abstract
      23531360 23531360
      Polycomb protein EZH2 regulates tumor invasion via the transcriptional repression of the metastasis suppressor RKIP in breast and prostate cancer.
      Gang Ren,Stavroula Baritaki,Himangi Marathe,Jingwei Feng,Sungdae Park,Sandy Beach,Peter S Bazeley,Anwar B Beshir,Gabriel Fenteany,Rohit Mehra,Stephanie Daignault,Fahd Al-Mulla,Evan Keller,Ben Bonavida,Ivana de la Serna,Kam C Yeung
      Cancer research  72  2012

      Show Abstract
      22505648 22505648
      Hypermethylated in cancer 1 (HIC1), a tumor suppressor gene epigenetically deregulated in hyperparathyroid tumors by histone H3 lysine modification.
      Svedlund, J; Koskinen Edblom, S; Marquez, VE; Åkerström, G; Björklund, P; Westin, G
      J Clin Endocrinol Metab  97  E1307-15  2012

      Show Abstract
      22544915 22544915
      Cooperation between Polycomb and androgen receptor during oncogenic transformation.
      Zhao, JC; Yu, J; Runkle, C; Wu, L; Hu, M; Wu, D; Liu, JS; Wang, Q; Qin, ZS; Yu, J
      Genome research  22  322-31  2012

      Show Abstract
      22179855 22179855

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      Advance your Epigenetics Research (MilliporeSigma)