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PE Mouse anti-EZH2
PE Mouse anti-EZH2
Flow cytometric analysis of EZH2 expression in a human cell line. Jurkat cells (ATCC, TIB-152™) were harvested, fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885) and stained with matching concentrations of either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse anti-EZH2 monoclonal antibody (Cat. No. 562478; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable Jurkat cells. Flow cytometry was performed using a BD FacsCanto™ II Flow Cytometer System.
Flow cytometric analysis of EZH2 expression in a human cell line. Jurkat cells (ATCC, TIB-152™) were harvested, fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885) and stained with matching concentrations of either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse anti-EZH2 monoclonal antibody (Cat. No. 562478; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable Jurkat cells. Flow cytometry was performed using a BD FacsCanto™ II Flow Cytometer System.
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BD Pharmingen™
Enhancer of zeste 2; ENX-1; Histone-lysine N-methyltransferase EZH2; KMT6
Human (QC Testing), Mouse, Rat, Dog, Chicken (Tested in Development)
Mouse IgG1
Human EZH2 recombinant protein aa. 156-256
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_11152951
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation and Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

推奨アッセイ手順

This antibody conjugate is suitable for intracellular staining of human cell lines using BD Cytofix™ Fixation Buffer (Cat. No. 554655). BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885) and BD Phosflow™ Perm Buffer III (Cat. No.558050) can  be used with this antibody conjugate.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  6. All other brands are trademarks of their respective owners.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562478 Rev. 1
抗体の詳細
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11/EZH2

The 11/EZH2 monoclonal antibody specifically binds to the methyltransferase, EZH2 (Enhancer of Zeste Homolog 2). EZH2 is a human homologue of Drosophila's Enhancer of zeste gene, an important regulator of homeobox gene expression. The EZH2 protein has a predicted molecular weight of ~85 kDa. EZH2 is a member of the Polycomb group (PcG) of proteins that are essential for the maintenance, but not initiation, of the transcriptionally repressed state of certain developmental genes. PcG proteins are a structurally diverse group of proteins with conserved functions from fly to human cells. PcG family proteins form multimeric complexes that regulate the expression of genes involved in cell cycle, DNA repair and differentiation. Specifically, EZH2 is a core enzymatic component of PRC2 (polycomb repressive complex 2). EZH2 is expressed in some lymph node follicular T cells and B cells. Thymocytes differentially express EZH2 at various stages during T-cell maturation. EZH2 interacts with multiple signaling proteins, including Vav, that are involved in lymphocyte development and activation. It is highly expressed in a variety of tumors including lymphomas as well as breast and prostate cancers. EZH2 is important in the self renewal and proliferation of numerous stem cell types including fetal hematopoietic stem cells, muscle satellite cells, hepatic stem/progenitor cells, neural stem cells, basal cell progenitors in the developing epidermis, embryonic stem cells, and some cancer stem cells.

  

562478 Rev. 1
フォーマットの詳細
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
562478 Rev.1
引用&参考文献
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Development References (10)

  1. Ezhkova E, Pasolli HA, Parker JS, et al. Ezh2 orchestrates gene expression for the stepwise differentiation of tissue-specific stem cells. Cell. 2009; 136(6):1122-1135. (Biology). View Reference
  2. Juan AH, Derfoul A, Feng X, et al. Polycomb EZH2 controls self-renewal and safeguards the transcriptional identity of skeletal muscle stem cells. Genes Dev. 2011; 25(8):789-794. (Biology). View Reference
  3. Kikuchi J, Kinoshita I, Shimizu Y, et al. Distinctive expression of the polycomb group proteins Bmi1 polycomb ring finger oncogene and enhancer of zeste homolog 2 in nonsmall cell lung cancers and their clinical and clinicopathologic significance. Cancer. 2010; 116(12):3015-3024. (Clone-specific: Immunohistochemistry, Western blot). View Reference
  4. Mochizuki-Kashio M, Mishima Y, Miyagi S, et al. Dependency on the polycomb gene Ezh2 distinguishes fetal from adult hematopoietic stem cells. Blood. 2011; 118(25):6553-6561. (Biology). View Reference
  5. Raaphorst FM, Otte AP, van Kemenade FJ. Distinct BMI-1 and EZH2 expression patterns in thymocytes and mature T cells suggest a role for Polycomb genes in human T cell differentiation. J Immunol. 2001; 166(10):5925-5934. (Biology). View Reference
  6. Shen X, Liu Y, Hsu YJ, et al. EZH1 mediates methylation on histone H3 lysine 27 and complements EZH2 in maintaining stem cell identity and executing pluripotency. Mol Cell. 2008; 32(4):491-502. (Biology). View Reference
  7. Simon JA, Lange CA. Roles of the EZH2 histone methyltransferase in cancer epigenetics. Mutat Res. 2008; 647(1-2):21-29. (Biology). View Reference
  8. Su IH, Dobenecker MW, Dickinson E, et al. Polycomb group protein ezh2 controls actin polymerization and cell signaling. Cell. 2005; 121(3):425-436. (Biology). View Reference
  9. Wolters T, Vissers KJ, Bangma CH, Schroder FH, van Leenders GJ. The value of EZH2, p27(kip1), BMI-1 and MIB-1 on biopsy specimens with low-risk prostate cancer in selecting men with significant prostate cancer at prostatectomy. BJU Int. 2009; 106(2):280-286. (Clone-specific: Immunohistochemistry). View Reference
  10. van Kemenade FJ, Raaphorst FM, Blokzijl T. Coexpression of BMI-1 and EZH2 polycomb-group proteins is associated with cycling cells and degree of malignancy in B-cell non-Hodgkin lymphoma. 2001; 97(12):3896-3901. (Biology). View Reference
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562478 Rev. 1

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