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APC Rat Anti-Mouse CD73
APC 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 APC Rat IgG1, κ Isotype Control (Cat. No. 554686; Left Plots) or APC Rat Anti-Mouse CD73 antibody (Cat. No. 567499; 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 APC Rat IgG1, κ Isotype Control (Cat. No. 554686; Left Plots) or APC Rat Anti-Mouse CD73 antibody (Cat. No. 567499; 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.

商品详情
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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


准备和存储

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 unconjugated antibody and free dye were removed.

推荐的实验流程

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.

商品通知

  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. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
567499 Rev. 1
抗体详情
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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.

        

567499 Rev. 1
格式详情
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC
Red 627-640 nm
651 nm
660 nm
567499 Rev.1
报价单和参考
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研发参考 (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). 查看参考
  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). 查看参考
  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). 查看参考
  4. Resta R, Yamashita Y, Thompson LF. Ecto-enzyme and signaling functions of lymphocyte CD73. Immunol Rev. 1998; 161:95-109. (Biology). 查看参考
  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). 查看参考
  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). 查看参考
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567499 Rev. 1

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