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APC Mouse Anti-Human DNAM-1 (CD226)
APC Mouse Anti-Human DNAM-1 (CD226)
Two-color flow cytometric analysis of DNAM-1 (CD226) expression on human peripheral blood leucocytes. Platelet-depleted human whole blood was stained with PE Mouse Anti-Human CD3 antibody (Cat. No. 555333) and with either APC Mouse IgG1, κ Isotype Control (Cat. No. 554681; Left Plot) or APC Mouse Anti-Human DNAM-1 (CD226) antibody (Cat. No. 567857/567858; 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.
Two-color flow cytometric analysis of DNAM-1 (CD226) expression on human peripheral blood leucocytes. Platelet-depleted human whole blood was stained with PE Mouse Anti-Human CD3 antibody (Cat. No. 555333) and with either APC Mouse IgG1, κ Isotype Control (Cat. No. 554681; Left Plot) or APC Mouse Anti-Human DNAM-1 (CD226) antibody (Cat. No. 567857/567858; 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.
Product Details
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BD Pharmingen™
DNAM-1; DNAM1; PTA1; TLiSA1
Human (QC Testing)
Mouse C57BL/6 IgG1, κ
Human NK Cells
Flow cytometry (Routinely Tested)
5 µl
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 to the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. 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.
  4. 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).
  5. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. An isotype control should be used at the same concentration as the antibody of interest.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567858 Rev. 1
Antibody Details
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11A8

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.

567858 Rev. 1
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC
Red 627-640 nm
651 nm
660 nm
567858 Rev.1
Citations & References
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Development References (7)

  1. 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
  2. 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
  3. 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
  4. 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.
  5. 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
  6. 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
  7. 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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567858 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products 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.

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.