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Multicolor flow cytometric analysis of CD150 (SLAM) expression by adult mouse spleen cells and bone marrow hematopoietic stem cells. Upper Plots: BALB/c spleen cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were stained with PE Rat Anti-Mouse CD45R/B220 (Cat. No. 553090/561878/553089) and either BD Horizon™ BV786 Rat IgG2a, κ Isotype Control (Cat. No. 563335; Left Plot) or BD Horizon™ BV786 Rat Anti-Mouse CD150 (SLAM) antibody (Cat. No. 567518; Right Plot) at 1 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. A bivariate pseudocolor density plot showing CD150 expression (or Ig Isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side-light scattering characteristics of viable (DAPI-negative) lymphocytes. Lower Plots: BALB/c mouse bone marrow cells were labeled with the BD IMag™ Mouse Hematopoietic Progenitor Enrichment Set (Cat. No. 558451) and separated on the BD IMag™ Cell Separation Magnet (Cat. No. 552311). The final hematopoietic progenitor-enriched fraction was subsequently stained with PerCP-Cy™5.5 Mouse Lineage Antibody Cocktail (Cat. No. 561317), PE Rat Anti-Mouse CD41 (Cat. No. 561850/558040), FITC Hamster Anti-Mouse CD48 (Cat. No. 557484) and BD Horizon™ BV786 Rat Anti-Mouse CD150 (SLAM) antibodies. The bivariate pseudocolor density plot showing the coexpressed levels of CD150 versus CD48 by viable (DAPI-negative) light scatter-gated cells (Left Plot) was further gated to show CD41 expression (Right Plot) on lineage-negative CD150+CD48- cells. Lineage-negative CD150+CD48-CD41- bone marrow cells have been reported to be highly enriched for adult mouse hematopoietic stem cells. Flow cytometry and data analysis was performed using a BD™ LSR II Flow Cytometer System and FloJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BV786 Rat Anti-Mouse CD150 (SLAM)
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 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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant™ Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant™ Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant™ Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant™ Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant™ Stain Buffer Plus (Cat. No. 566385).
Product Notices
- 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.
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Companion Products
The Q38-480 monoclonal antibody specifically binds to mouse CD150, also known as SLAM (signaling lymphocyte activation molecule). CD150 is a type 1 transmembrane glycoprotein that is a member of the CD2 subfamily of the Ig superfamily. It is encoded by the Slamf1 (signaling lymphocytic activation molecule family member 1) gene. CD150 is differentially expressed on subsets of thymocytes, T and B lymphocytes, dendritic cells, macrophages, and endothelial cells. SLAM plays multiple roles in innate and adaptive immunity serving as an adhesion molecule and/or coreceptor. CD150-mediated costimulation of TCR-activated T cells reportedly results in the increased production of IFN-γ by Th1 cells and is required for IL-4 production by T follicular helper cells. CD150 also plays important roles in hematopoietic cell developmental pathways. CD150 is differentially expressed by self-renewing adult hematopoietic stem cells (HSC) whereas non-multipotent hematopoietic progenitor cells are CD150-. Utilizing additional cell surface markers, lineage-negative CD150+CD48-CD41- cell fractions are reported to be highly enriched for adult HSC.
The antibody was conjugated to BD Horizon™ BV786 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon™ BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 786-nm. BD Horizon™ BV786 can be excited by the violet laser and detected in a filter used to detect Cy™7-like dyes (eg, 780/60-nm filter).
Development References (5)
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Castro AG, Hauser TM, Cocks BG, et al. Molecular and functional characterization of mouse signaling lymphocytic activation molecule (SLAM): differential expression and responsiveness in Th1 and Th2 cells.. J Immunol. 1999; 163(11):5860-70. (Biology). View Reference
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Guo H, Ma O, Friedman AD. The Cebpa +37-kb enhancer directs transgene expression to myeloid progenitors and to long-term hematopoietic stem cells.. J Leukoc Biol. 2014; 96(3):419-26. (Clone-specific: Flow cytometry). View Reference
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Kiel MJ, Yilmaz OH, Iwashita T, Terhorst C, Morrison SJ. SLAM family receptors distinguish hematopoietic stem and progenitor cells and reveal endothelial niches for stem cells. Cell. 2005; 121(7):1109-1121. (Biology). View Reference
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Wang N, Satoskar A, Faubion W, et al. The cell surface receptor SLAM controls T cell and macrophage functions. J Exp Med. 2004; 199(9):1255-1264. (Biology). View Reference
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Yusuf I, Kageyama R, Monticelli L, et al. Germinal center T follicular helper cell IL-4 production is dependent on signaling lymphocytic activation molecule receptor (CD150). J Immunol. 2010; 185(1):190-202. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.