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PE Rat Anti-Mouse CD226 (DNAM-1)
PE Rat Anti-Mouse CD226 (DNAM-1)

Two-color flow cytometric analysis of CD226 expression on mouse splenic leucocytes. C57BL/6 mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD8a antibody (Cat. No. 553035/561093) and either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plot) or PE Rat Anti-Mouse CD226 (DNAM-1) antibody (Cat. No. 567357; Right Plot) at 0.25 μg/test. Bivariate pseudocolor density plots showing the correlated expression of CD226 (or Ig Isotype control staining) versus CD8a were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System equipped with a Yellow-Green Laser. Data shown on this Technical Data Sheet are not lot specific.

Two-color flow cytometric analysis of CD226 expression on mouse splenic leucocytes. C57BL/6 mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD8a antibody (Cat. No. 553035/561093) and either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plot) or PE Rat Anti-Mouse CD226 (DNAM-1) antibody (Cat. No. 567357; Right Plot) at 0.25 μg/test. Bivariate pseudocolor density plots showing the correlated expression of CD226 (or Ig Isotype control staining) versus CD8a were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System equipped with a Yellow-Green Laser. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Pharmingen™
CD226 antigen; Cd226; DNAM-1; DNAM1; Pta1; TLiSA1
Mouse (QC Testing)
Rat IgG2a, κ
Mouse DNAM-1 transfected cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing ≤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 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567357 Rev. 1
Antibody Details
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TX42.1

The TX42.1 monoclonal antibody specifically recognizes 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 (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 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.

567357 Rev. 1
Format Details
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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.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
567357 Rev.1
Citations & References
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Development References (5)

  1. Shibuya K, Shibata K, Tahara-Hanaoka S, Shibuya A. Comment on "CD226 is specifically expressed on the surface of Th1 cells and regulates their expansion and effector functions".. J Immunol. 2006; 176(7):3855; author reply 3856. (Clone-specific: Flow cytometry). View Reference
  2. Stanietsky N, Rovis TL, Glasner A, et al. Mouse TIGIT inhibits NK-cell cytotoxicity upon interaction with PVR.. Eur J Immunol. 2013; 43(8):2138-50. (Clone-specific: Flow cytometry). View Reference
  3. Tahara-Hanaoka S, Miyamoto A, Hara A, Honda S, Shibuya K, Shibuya A. Identification and characterization of murine DNAM-1 (CD226) and its poliovirus receptor family ligands.. Biochem Biophys Res Commun. 2005; 329(3):996-1000. (Immunogen: Blocking, Flow cytometry, Functional assay, Inhibition). View Reference
  4. Tahara-Hanaoka S, Shibuya K, Kai H, et al. Blood. 2006; 107(4):491-496. (Clone-specific: Blocking, Flow cytometry, Functional assay, In vivo exacerbation, Stimulation). View Reference
  5. Yamashita Y, Abe F, Hirochika R, Tahara-Hanaoka S, Shibuya A, Shibuya K. Establishment and characterization of a novel anti-DNAM-1 monoclonal antibody.. Monoclon Antib Immunodiagn Immunother. 2013; 32(1):60-4. (Clone-specific: In vivo exacerbation). View Reference
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567357 Rev. 1

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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.