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APC Mouse Anti-Human CD215 (IL-15Rα)
APC Mouse Anti-Human CD215 (IL-15Rα)
Multiparameter flow cytometric analysis of CD215 (IL15Rα) expression on human monocytes. Human peripheral blood mononuclear cells were cultured in complete tissue culture medium without (Left Plots) or with (Right Plots) lipopolysaccharide (LPS, 100 ng/ml) and Recombinant Human IFN-γ protein (Cat. No. 554617, 10 ng/ml) for 20 hours at 37°C. The cells were harvested, preincubated with Human BD Fc Block™ (Cat. No. 564219), and then stained with BD Horizon™ BV421 Mouse Anti-Human CD14 antibody (Cat. No. 565283) and with either APC Mouse IgG2b, κ Isotype Control (Cat. No. 565381; Upper Plots) or APC Mouse Anti-Human CD215 (IL15Rα) antibody (Cat. No. 567740/567741; Lower Plots).  BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. Bivariate pseudocolor density plots showing the correlated expression of CD215 (IL15Rα) [or Ig Isotype control staining] versus CD14 were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) monocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of CD215 (IL15Rα) expression on human monocytes. Human peripheral blood mononuclear cells were cultured in complete tissue culture medium without (Left Plots) or with (Right Plots) lipopolysaccharide (LPS, 100 ng/ml) and Recombinant Human IFN-γ protein (Cat. No. 554617, 10 ng/ml) for 20 hours at 37°C. The cells were harvested, preincubated with Human BD Fc Block™ (Cat. No. 564219), and then stained with BD Horizon™ BV421 Mouse Anti-Human CD14 antibody (Cat. No. 565283) and with either APC Mouse IgG2b, κ Isotype Control (Cat. No. 565381; Upper Plots) or APC Mouse Anti-Human CD215 (IL15Rα) antibody (Cat. No. 567740/567741; Lower Plots).  BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. Bivariate pseudocolor density plots showing the correlated expression of CD215 (IL15Rα) [or Ig Isotype control staining] versus CD14 were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) monocytes. 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™
IL15RA; IL-15R-alpha; IL-15RA; IL-15Rα; CD215
Human (QC Testing)
Mouse BALB/c IgG2b, κ
Recombinant Human IL-15Rα Extracellular Domain
Flow cytometry (Routinely Tested)
5 µl
IX 234
3601
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 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® 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).
Antibody Details
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JM7A4

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.

        

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
Citations & References
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Development References (5)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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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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.