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Flow cytometric analysis using BD OptiBuild™ BV421 Mouse Anti-Human CD215 (IL-15Rα) antibody (Cat. No. 747704; solid line histogram) on monocytes stimulated with Lipopolysaccharide and Recombinant Human IFN-γ for 20 hours with Isotype Control (dotted line histogram) Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
BD OptiBuild™ BV421 Mouse Anti-Human CD215 (IL-15Rα)
Regulatory Status Legend
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
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
Product Notices
- This antibody was developed for use in flow cytometry.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
Companion Products
The JM7A4 monoclonal antibody specifically binds to CD215, which is also known as the IL-15 Receptor alpha subunit (IL-15R-alpha, IL-15Ra, or IL-15Rα). This type I transmembrane glycoprotein is encoded by IL15RA (Interleukin 15 receptor subunit alpha) and is variably expressed on macrophages, natural killer (NK) cells, T cells and B cells and by some nonlymphoid cells including fibroblasts. Although it can independently bind IL-15 with high affinity, it does not contain a signaling motif. CD215 (IL-15Rα) can present IL-15 in cis or trans fashion to the IL-2/15R beta (CD122) and IL-2R gamma (γc or CD132) receptor complex which can then transduce signals intracellularly. Several different CD215 (IL-15Rα) isoforms have been described that are produced by alternative splicing and may alter signal transduction responses to IL-15. A cleaved soluble form of CD215 known as sIL-15RA has also been reported which can bind and antagonize IL-15 activity. By binding to its heterotrimeric receptor, IL-15 plays crucial roles in innate immunity, eg, through the activation of NK cells and adaptive immunity, eg, in enhancing the survival of CD8+ memory T cells.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Development References (5)
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Dubois S, Mariner J, Waldmann TA, Tagaya Y. IL-15Ralpha recycles and presents IL-15 In trans to neighboring cells.. Immunity. 2002; 17(5):537-47. (Immunogen: Flow cytometry). View Reference
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Llinas L, Lazaro A, de Salort J, Matesanz-Isabel J, Sintes J, Engel P. Expression profiles of novel cell surface molecules on B-cell subsets and plasma cells as analyzed by flow cytometry. Immunol Lett. 2011; 134(2):113-121. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
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Matesanz-Isabel J, Sintes J, Llinas L, de Salort J, Lazaro A, Engel P. New B-cell CD molecules. Immunol Lett. 2011; 134(2):104-112. (Clone-specific: Flow cytometry). View Reference
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Mortier E, Bernard J, Plet A, Jacques Y. Natural, proteolytic release of a soluble form of human IL-15 receptor alpha-chain that behaves as a specific, high affinity IL-15 antagonist.. J Immunol. 2004; 173(3):1681-8. (Biology). View Reference
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Vámosi G, Bodnár A, Vereb G, et al. IL-2 and IL-15 receptor alpha-subunits are coexpressed in a supramolecular receptor cluster in lipid rafts of T cells.. Proc Natl Acad Sci USA. 2004; 101(30):11082-7. (Clone-specific: Immunofluorescence). View Reference
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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.