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PE Rat Anti-Mouse CD73
PE Rat Anti-Mouse CD73
Two-color flow cytometric analysis of CD73 expression on splenic leucocytes from two mouse strains. C57BL/6 (Top Plots) and BALB/C (Bottom Plots) mouse splenic leucocytes were stained with APC Armenian Hamster Anti-Mouse CD3e antibody (Cat. No. 553066/561826) and either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Left Plots) or PE Rat Anti-Mouse CD73 (Cat. No. 567215; Right Plots) at 0.25 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The pseudocolor density plots showing the correlated expression of CD73 (or Ig Isotype control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of CD73 expression on splenic leucocytes from two mouse strains. C57BL/6 (Top Plots) and BALB/C (Bottom Plots) mouse splenic leucocytes were stained with APC Armenian Hamster Anti-Mouse CD3e antibody (Cat. No. 553066/561826) and either PE Rat IgG1, κ Isotype Control (Cat. No. 553925; Left Plots) or PE Rat Anti-Mouse CD73 (Cat. No. 567215; Right Plots) at 0.25 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The pseudocolor density plots showing the correlated expression of CD73 (or Ig Isotype control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
5'-NT; 5'-nucleotidase; Nt5e; ecto-5'-nucleotidase
Mouse (QC Testing)
Rat WI, also known as Wistar (outbred) IgG1, κ
Mouse CD73-transfected CHO Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
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 with R-PE under optimum conditions, and unconjugated antibody and free PE 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 Comp beads 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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567215 Rev. 2
Antibody Details
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TY/11.8

The TY/11.8 monoclonal antibody specifically recognizes mouse CD73 which is also known as Ecto-5'-nucleotidase (5'-NT). CD73 is a ~69 kDa glycosylphosphatidylinositol (GPI)-anchored, cell-surface glycoprotein that is encoded by Nt5e which belongs to the 5'-nucleosidase family. CD73 expression appears to be developmentally regulated on leucocytes. In the bone marrow, it is found on most CD11b+ myeloid cells and very few CD19+ cells of the B-lymphocyte lineage. It is neither found expressed on CD11b+ cells in the periphery nor on bone marrow-derived GM-CSF-stimulated dendritic cells. Some peripheral B lymphocytes express CD73, with higher levels detected on Ig isotype-switched B cells. The few thymocytes which have detectable surface CD73 are found within CD4-CD8-and the CD4+CD8- subpopulations. In peripheral lymphoid organs, large proportions of the CD4+ and CD8+ T lymphocytes express CD73. Significant variation in the frequencies of peripheral CD73+ T cells have been observed amongst inbred mouse strains. For example, C57BL/6 mice reportedly have higher frequencies of peripheral CD73+ T cells when compared with BALB/c mice. In the thymus and peripheral lymphoid organs, CD73 is found on endothelial and stromal cells. CD73 has also been detected on bone marrow and thymic epithelial cell lines, kidney glomeruli and proximal-tubule epithelial cells, liver endothelial cells and hepatocytes, mesenchymal cells, and fibroblasts including cancer-associated fibroblasts (CAFs) found in tumors. CD73 has enzymatic and signal transduction activities. It catalyzes the dephosphorylation of extracellular nucleoside 5' monophosphates to nucleosides. CD73 acts on adenosine monophosphate (AMP) to generate and regulate the concentration of extracellular adenosine. Adenosine can bind to adenosine receptors expressed on cells in many tissues and regulate physiological responses including anti-inflammatory or immunosuppressive responses. Regulatory T cells (Treg) can generate immunosuppressive adenosine by their expression and activity of the CD39 and CD73 ectoenzymes.

        

567215 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
567215 Rev.2
Citations & References
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View product citations for antibody "567215" on CiteAb

Development References (6)

  1. Barron L, Dooms H, Hoyer KK, et al. Cutting edge: mechanisms of IL-2-dependent maintenance of functional regulatory T cells.. J Immunol. 2010; 185(11):6426-30. (Clone-specific: Flow cytometry). View Reference
  2. Deaglio S, Dwyer KM, Gao W, et al. Adenosine generation catalyzed by CD39 and CD73 expressed on regulatory T cells mediates immune suppression.. J Exp Med. 2007; 204(6):1257-65. (Biology). View Reference
  3. Le Texier L, Lineburg KE, Cao B, et al. Autophagy-dependent regulatory T cells are critical for the control of graft-versus-host disease. JCI Insight. 2016; 1(15):e86850. (Clone-specific: Flow cytometry). View Reference
  4. Resta R, Yamashita Y, Thompson LF. Ecto-enzyme and signaling functions of lymphocyte CD73. Immunol Rev. 1998; 161:95-109. (Biology). View Reference
  5. Yamashita Y, Hooker SW, Jiang H, et al. CD73 expression and fyn-dependent signaling on murine lymphocytes. Eur J Immunol. 1998; 28(10):2981-2990. (Immunogen: Flow cytometry). View Reference
  6. Yu M, Guo G, Huang L, et al. CD73 on cancer-associated fibroblasts enhanced by the A(2B)-mediated feedforward circuit enforces an immune checkpoint. Nature Commun. 2020; 11(1):515. (Clone-specific: Flow cytometry). View Reference
View All (6) View Less
567215 Rev. 2

 

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