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R718 Rat Anti-Mouse CD150 (SLAM)
R718 Rat Anti-Mouse CD150 (SLAM)

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. 553090/553089/561878) and either BD Horizon™ R718 Rat IgG2a, κ Isotype Control (Cat. No. 566941; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse CD150 (SLAM) antibody (Cat. No. 567735; 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. 558040/561850), FITC Hamster Anti-Mouse CD48 (Cat. No. 557484) and BD Horizon™ R718 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) were 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.

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. 553090/553089/561878) and either BD Horizon™ R718 Rat IgG2a, κ Isotype Control (Cat. No. 566941; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse CD150 (SLAM) antibody (Cat. No. 567735; 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. 558040/561850), FITC Hamster Anti-Mouse CD48 (Cat. No. 557484) and BD Horizon™ R718 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) were 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.

Product Details
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BD Horizon™
SLAM; Slamf1; Signaling lymphocytic activation molecule family member 1
Mouse (QC Testing)
Rat IgG2a, κ
Mouse CD150 Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
27218
Aqueous buffered solution containing ≤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 unreacted dye was removed.

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.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Alexa Fluor™ is a trademark of Life Technologies Corporation.
567735 Rev. 1
Antibody Details
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Q38-480

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™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon™ Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.

567735 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
R718
Red 627-640 nm
695 nm
718 nm
567735 Rev.1
Citations & References
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Development References (5)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
View All (5) View Less
567735 Rev. 1

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.