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R718 Rat Anti-Mouse CD73
R718 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 preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and stained with  BD Horizon™ BUV395 Hamster Anti-Mouse CD3e antibody  (Cat.  No. 563565/565992)  and  with either  BD Horizon™ R718 Rat  IgG1,  κ  Isotype  Control  (Cat.  No. 566948;  Left  Plots)  or  BD Horizon™  R718 Rat  Anti-Mouse  CD73 antibody (Cat.  No.  567644; Right  Plots)  at  0.5  μg/test.  DAPI 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™  X-20 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 preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and stained with  BD Horizon™ BUV395 Hamster Anti-Mouse CD3e antibody  (Cat.  No. 563565/565992)  and  with either  BD Horizon™ R718 Rat  IgG1,  κ  Isotype  Control  (Cat.  No. 566948;  Left  Plots)  or  BD Horizon™  R718 Rat  Anti-Mouse  CD73 antibody (Cat.  No.  567644; Right  Plots)  at  0.5  μg/test.  DAPI 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™  X-20 Cell  Analyzer  System  and  FlowJo™  software.  Data shown  on  this  Technical  Data  Sheet  are  not  lot  specific.

製品詳細
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BD Horizon™
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 to the dye under optimum conditions and unreacted dye was removed.

推奨アッセイ手順

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 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. 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.
  3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  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.
567644 Rev. 1
抗体の詳細
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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.

        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.

567644 Rev. 1
フォーマットの詳細
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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.
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R718
Red 627-640 nm
695 nm
718 nm
567644 Rev.1
引用&参考文献
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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
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567644 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.