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Flow cytometric analysis of CD226 expression on human peripheral blood lymphocytes. Human whole blood was depleted of platelets, washed, and preincubated with 5% normal human serum. The cells were then stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human CD226 antibody (Cat. No. 564797; solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histogram showing CD226 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
BD Pharmingen™ Alexa Fluor® 647 Mouse Anti-Human CD226
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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
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Companion Products
The DX11 monoclonal antibody specifically binds to CD226 which is also known as DNAX accessory molecule-1 (DNAM-1), Platelet and T cell activation antigen 1 (PTA1), or T lineage-specific activation antigen 1 antigen (TLiSA1). CD226 is a 65 kDa type 1 transmembrane glycoprotein consisting of 318 amino acid residues including two Ig-like domains. CD226 is expressed on the majority of T cells, NK cells, monocytes, platelets, and a subset of B cells, but not on erythrocytes. It is also present on a subset of thymocytes coexpressing high density surface CD3. CD226 is not present on normal fibroblast cell lines or tumor cell lines of epithelial or neuronal origins. CD226 is a tyrosine phosphorylated, signal-transducing molecule which participates in primary adhesion during cytotoxic T lymphocyte (CTL)- or NK cell-mediated cytotoxicity. The DX11 antibody inhibits T- and NK cell-mediated cytotoxicity against a variety of tumor cell targets, and blocks cytokine production by alloantigen-specific T cells.
Development References (4)
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Lanier LL, Shibuya A, Burns G. CD226 (DNAM-1, PTA1, Tlisa). In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:921-922.
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Shibuya A, Campbell D, Hannum C, et al. DNAM-1, a novel adhesion molecule involved in the cytolytic function of T lymphocytes. Immunity. 1996; 4(6):573-581. (Immunogen: Bioassay, Blocking, Flow cytometry, Functional assay, Immunoaffinity chromatography, Inhibition, Western blot). View Reference
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Shibuya A, Lanier LL, Phillips JH. Protein kinase C is involved in the regulation of both signaling and adhesion mediated by DNAX accessory molecule-1 receptor. J Immunol. 1998; 161(4):1671-1676. (Clone-specific: Bioassay, Blocking, Cytotoxicity, Flow cytometry, Functional assay, Immunoprecipitation). View Reference
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Zola H, Swart B, Boumsell L, Mason DY. Human Leucocyte Differentiation Antigen nomenclature: update on CD nomenclature. Report of IUIS/WHO Subcommittee.. J Immunol Methods. 2003; 275(1-2):1-8. (Clone-specific: Flow cytometry). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.