Alexa Fluor® 647 Mouse Anti-EZH2
Clone 11/EZH2 (RUO)
- Brand BD Pharmingen™
- Alternative Name Enhancer of zeste 2; ENX-1; Histone-lysine N-methyltransferase EZH2; KMT6
- Vol. Per Test 5 µl
- Isotype Mouse IgG1
- Reactivity Human (QC Testing) Mouse, Rat, Chicken, Dog (Tested in Development)
Intracellular staining (flow cytometry) (Routinely Tested)
Bioimaging, Immunofluorescence (Tested During Development)
- Immunogen Human EZH2 recombinant protein aa. 156-256
- Storage Buffer Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
- 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).
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Species testing during development may have been performed with a different format of the same clone. Selected applications have been tested for cross-reactivity.
- An isotype control should be used at the same concentration as the antibody of interest.
- Triton is a trademark of the Dow Chemical Company.
· This antibody conjugate is suitable for intracellular staining of human cell lines using BD Cytofix™ Fixation Buffer. 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.
· The Bioimaging protocol can be found at http://www.bdbiosciences.com/support/resources/protocols/ceritifed_reagents.jsp.