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Two-color flow cytometric analysis of DNAM-1 (CD226) expression on human peripheral blood leucocytes. Platelet-depleted human whole blood was stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 555335) and with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon™ Mouse Anti-Human DNAM-1 (CD226) antibody (Cat. No. 568071/568072; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of DNAM-1 (CD226) [or Ig Isotype control staining] versus CD3 was derived from gated events based on the forward and side-light scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
BD Horizon™ BV421 Mouse Anti-Human DNAM-1 (CD226)
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For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant™ Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant™ Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant™ Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant™ Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant™ Stain Buffer Plus (Cat. No. 566385).
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The 11A8 monoclonal antibody specifically recognizes DNAX accessory molecule-1 (DNAM-1) which is also known as CD226, Platelet and T cell activation antigen 1 (PTA1), or T lineage-specific activation antigen 1 antigen (TLiSA1). CD226 is an ~65 kDa type I transmembrane glycoprotein that is encoded by CD226 (CD226 antigen) which belongs to the immunoglobulin superfamily (IgSF). The N-terminal extracellular region of CD226 (DNAM-1) contains two C2 type Ig-like domains which are followed by a transmembrane region and a cytoplasmic tail with predicted phosphorylation sites involved in signaling. This adhesion molecule is expressed on subsets of thymocytes, peripheral CD4+ and CD8+ T cells and B cells, NK cells, monocytes, macrophages, and platelets. CD226 (DNAM-1) associates with leukocyte function associated antigen-1 (LFA-1) and is involved in TCR-mediated signal transduction for T cell proliferation and differentiation. Interactions between the CD226 (DNAM-1) activating receptor and its ligands, CD112 and CD155, results in cellular signaling that promotes innate and adaptive immune responses, including the activation, differentiation, and survival of cytotoxic cells.
Development References (7)
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Cella M, Presti R, Vermi W, et al. Loss of DNAM-1 contributes to CD8+ T-cell exhaustion in chronic HIV-1 infection.. Eur J Immunol. 2010; 40(4):949-54. (Clone-specific: Flow cytometry, Functional assay). View Reference
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Fourcade J, Sun Z, Chauvin JM, et al. CD226 opposes TIGIT to disrupt Tregs in melanoma.. JCI Insight. 2018; 3(14):121157. (Clone-specific: Flow cytometry). View Reference
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Fuchs A, Cella M, Giurisato E, Shaw AS, Colonna M. Cutting edge: CD96 (tactile) promotes NK cell-target cell adhesion by interacting with the poliovirus receptor (CD155).. J Immunol. 2004; 172(7):3994-8. (Immunogen: Blocking, Functional assay, Stimulation). View Reference
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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. (Biology). 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. (Biology). 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. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.
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.