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Multicolor flow cytometric analysis of CD215 (IL-15Rα) expression on mature and immature bone marrow-derived dendritic cells. Mouse bone marrow-derived dendritic cells were either not stimulated (Left Figures) or stimulated (Middle Figures) by LPS overnight (37°C). The cells were harvested, washed and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Hamster Anti-Mouse CD11c antibody (Cat. No. 561119) and either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Top Figures) or PE Rat Anti-Mouse CD215 (IL-15Rα) antibody (Cat. No. 568235; Bottom Figures) at 0.125 µg/test. 7-AAD (7-Amino-Actinomycin D) Solution (Cat. No. 559925) was added to cells right before analysis. Bivariate pseudocolor density plots showing the correlated expression of CD215 (IL-15Rα) (or Ig Isotype control staining) versus CD11c were derived from gated events with the forward and side light- scatter characteristics of viable (7-AAD-negative) cells. Bone marrow-derived dendritic cells were identified as CD11c-positive cells. Histograms (Right Figure) were derived from gated cell subset show CD215 level for Isotype Control (dashed line histogram) and dendritic cells (solid line histogram). Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ PE Rat Anti-Mouse 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
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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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 http://regdocs.bd.com to access safety data sheets (SDS).
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
The monoclonal antibody W11-873 specifically reacts with mouse CD215, which is also known as IL-15 Receptor Alpha, IL-15Ra, or IL-15Rα. CD215 is the high-affinity alpha subunit of the interleukin-15 receptor. CD215 is structurally similar to CD25 (IL-2Rα) and forms a heterotrimer with CD122 (IL-2Rβ) and CD132 (IL-2Rγ/common gamma chain). CD215 can bind to IL-15 with high affinity even in the absence of CD122/CD132. The expression of CD215 has been detected in various immune cells such as T cells, B cells, dendritic cells, macrophages, and NK cells, as well as non-immune cell types. CD215 has both a membrane-bound form and a soluble form generated from proteolytic cleavage. IL-15-mediated signaling activates the JAK/STAT pathway and modulates both innate and adaptive immune responses. IL-15 signaling is critical for the long-term maintenance of memory CD8+ T cells and their ability to mount a secondary response upon rechallenge.
Development References (7)
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Anderson DM, Kumaki S, Ahdieh M, et al. Functional characterization of the human interleukin-15 receptor alpha chain and close linkage of IL15RA and IL2RA genes.. J Biol Chem. 1995; 270(50):29862-9. (Biology). View Reference
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Budagian V, Bulanova E, Paus R, Bulfone-Paus S. IL-15/IL-15 receptor biology: a guided tour through an expanding universe.. Cytokine Growth Factor Rev. 2006; 17(4):259-80. (Biology). View Reference
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Giri JG, Kumaki S, Ahdieh M, et al. Identification and cloning of a novel IL-15 binding protein that is structurally related to the alpha chain of the IL-2 receptor.. EMBO J. 1995; 14(15):3654-63. (Biology). View Reference
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Giron-Michel J, Giuliani M, Fogli M, et al. Membrane-bound and soluble IL-15/IL-15Ralpha complexes display differential signaling and functions on human hematopoietic progenitors.. Blood. 2005; 106(7):2302-10. (Biology). View Reference
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Lorenzen I, Dingley AJ, Jacques Y, Grötzinger J. The structure of the interleukin-15 alpha receptor and its implications for ligand binding.. J Biol Chem. 2006; 281(10):6642-7. (Biology). View Reference
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Richer MJ, Pewe LL, Hancox LS, Hartwig SM, Varga SM, Harty JT. Inflammatory IL-15 is required for optimal memory T cell responses.. J Clin Invest. 2015; 125(9):3477-90. (Biology). View Reference
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Sato N, Patel HJ, Waldmann TA, Tagaya Y. The IL-15/IL-15Ralpha on cell surfaces enables sustained IL-15 activity and contributes to the long survival of CD8 memory T cells.. Proc Natl Acad Sci U S A. 2007; 104(2):588-93. (Biology). View Reference
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