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Preparation And Storage
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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- An isotype control should be used at the same concentration as the antibody of interest.
- PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
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The 10E5 monoclonal antibody specifically binds to CD226 which is also known as, Platelet and T-cell activation antigen 1 (Pta1), T lineage-specific activation antigen 1 (TLiSA1), or DNAX accessory molecule-1 (DNAM-1). CD226 is a type I transmembrane glycoprotein that belongs to the immunoglobulin superfamily. CD226 serves as an adhesion molecule that is primarily expressed on T lymphocytes. Naïve CD8+ T cells express high levels of CD226 whereas it is differentially expressed on CD4+ T cells. In certain model systems, long term-cloned Th1 cells can express high CD226 levels when compared with the downregulated CD226 expression exhibited on cloned Th2 and Th0 cells. CD226 expression is also detectable on subsets of CD11b+ macrophages and NK cells. Adhesive interactions between CD226 and its ligands, CD112 and CD155, can result in cell signaling events that promote innate and adaptive immune responses, including the differentiation and survival of cytotoxic cells.
Development References (4)
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Dardalhon V, Schubart AS, Reddy J, et al. CD226 is specifically expressed on the surface of Th1 cells and regulates their expansion and effector functions. Immunology. 2005; 175(3):1558-1565. (Immunogen: Flow cytometry, Inhibition, In vivo exacerbation). View Reference
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Seth S, Georgoudaki AM, Chambers BJ et al. Heterogeneous expression of the adhesion receptor CD226 on murine NK and T cells and its function in NK-mediated killing of immature dendritic cells. J Leukoc Biol. 2009; 86(1):91-101. (Biology). View Reference
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Tahara-Hanaoka S, Shibuya K, Kai H, et al. Blood. 2006; 107(4):491-496. (Biology). View Reference
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Tahara-Hanaoka S, Shibuya K, Onoda Y et al. Functional characterization of DNAM-1 (CD226) interaction with its ligands PVR (CD155) and nectin-2 (PRR-2/CD112). Int Immunol. 2006; 16(4):533-538. (Biology). View Reference
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.