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PerCP-Cy5.5 Mouse Anti-Human CD39
PerCP-Cy5.5 Mouse Anti-Human CD39

Flow cytometric analysis of CD39 expression on human peripheral blood leucocyte populations.

Upper Plots: Whole blood was stained with either PerCP-Cy5.5 Mouse IgG1, κ Isotype Control (Cat No. 550795; Left Plot) or PerCP-Cy5.5 Mouse Anti-Human CD39 antibody (Cat No. 567534/567535; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD39 (or Ig Isotype control staining) versus side light-scatter signals (SSC-A) was derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations.

Lower Plots: Human peripheral blood mononuclear cells (PBMC) were preincubated with Human BD Fc Block™ (Cat. No. 564219/564220) and then stained with BD Horizon™ BUV395 Mouse Anti-Human CD4 (Cat. No. 564724), FITC Mouse Anti-Human CD25 (Cat. No. 555431/560990), BD Horizon™ BUV737 Mouse Anti-Human CD127 (Cat No. 612795/612794) antibodies, and either PerCP-Cy5.5 Mouse IgG1, κ Isotype Control (dashed line histogram) or PerCP-Cy5.5 Mouse Anti-Human CD39 antibody (solid line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the coexpressed levels of CD25 versus CD127 by viable (DAPI-negative) light scatter-gated CD4+ T cells [Left Plot] was further gated to reveal CD39 expression (or Ig Isotype control staining) [Right Plot] on CD4+CD25+CD127low T cells (ie, cells with a Regulatory T cell immunophenotype) as shown.

Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software.

Flow cytometric analysis of CD39 expression on human peripheral blood leucocyte populations.

Upper Plots: Whole blood was stained with either PerCP-Cy5.5 Mouse IgG1, κ Isotype Control (Cat No. 550795; Left Plot) or PerCP-Cy5.5 Mouse Anti-Human CD39 antibody (Cat No. 567534/567535; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD39 (or Ig Isotype control staining) versus side light-scatter signals (SSC-A) was derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations.

Lower Plots: Human peripheral blood mononuclear cells (PBMC) were preincubated with Human BD Fc Block™ (Cat. No. 564219/564220) and then stained with BD Horizon™ BUV395 Mouse Anti-Human CD4 (Cat. No. 564724), FITC Mouse Anti-Human CD25 (Cat. No. 555431/560990), BD Horizon™ BUV737 Mouse Anti-Human CD127 (Cat No. 612795/612794) antibodies, and either PerCP-Cy5.5 Mouse IgG1, κ Isotype Control (dashed line histogram) or PerCP-Cy5.5 Mouse Anti-Human CD39 antibody (solid line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the coexpressed levels of CD25 versus CD127 by viable (DAPI-negative) light scatter-gated CD4+ T cells [Left Plot] was further gated to reveal CD39 expression (or Ig Isotype control staining) [Right Plot] on CD4+CD25+CD127low T cells (ie, cells with a Regulatory T cell immunophenotype) as shown.

Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software.

Product Details
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BD Pharmingen™
A1; ENTPD1; NTPDase-1; ecto-ATPDase 1; ecto-ATPase 1; ecto-apyrase
Human (QC Testing)
Mouse BALB/c IgG1, κ
PHA-activated Human Peripheral Blood Mononuclear Cells
Flow cytometry (Routinely Tested)
5 µl
V CD39.07; VI CD39,1
Aqueous buffered solution containing BSA and ≤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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

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. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  6. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  7. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  8. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  11. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567534 Rev. 1
Antibody Details
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A1

The A1 monoclonal antibody specifically recognizes human CD39 which is also known as Ecto-ATP diphosphohydrolase 1 (Ecto-ATPase 1 or Ecto-ATPDase 1), Ecto-apyrase or NTPDase 1. CD39 is a ~78 kDa integral membrane glycoprotein that is encoded by ENTPD1 (Ectonucleoside triphosphate diphosphohydrolase 1). CD39 contains two transmembrane domains, one having a N- and the other a C-terminal cytoplasmic tail, and a large extracellular domain that has the enzymatic site. CD39 is also known as Lymphoid cell activation antigen because its expression is induced upon activation of T and B cells. CD39 is variably expressed on some regulatory T cells, NK cells, granulocytes, monocytes, dendritic cells, Langerhans cells, endothelial cells, platelets, and neurons. CD39 is a member of the ectonucleoside triphosphate dihydrolases (E-NTPDases) family that is involved in the regulation of extracellular nucleotide catabolism by controlling the extracellular nucleoside triphosphate pool (NTP). It functions as an ectoenzyme that can hydrolyze both nucleoside triphosphates and diphosphates such as ATP and ADP and thereby suppress inflammation and regulate platelet activation as well as purinergic neurotransmission. The ectoenzymes CD39 and CD73 can act in tandem to enable regulatory T cells (Treg) to generate immunosuppressive adenosine and thereby regulate immune responses.

567534 Rev. 1
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
567534 Rev.1
Citations & References
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Development References (6)

  1. Allard B, Longhi MS, Robson SC, Stagg J. The ectonucleotidases CD39 and CD73: Novel checkpoint inhibitor targets.. Immunol Rev. 2017; 276(1):121-144. (Biology). View Reference
  2. Aversa GG, Suranyi MG, Waugh JA, Bishop AG, Hall BM. Detection of a late lymphocyte activation marker by A1, a new monoclonal antibody.. Transplant Proc. 1988; 20(1):49-52. (Immunogen: Flow cytometry). View Reference
  3. Aversa GG, Waugh JA, Bishop GA, Hall BM. Use of monoclonal antibodies to study in vivo and in vitro-activated lymphocytes.. Transplant Proc. 1989; 21(1 Pt 1):349-50. (Clone-specific: Flow cytometry). View Reference
  4. Gouttefangeas C, Mansur I, Bensussan A, Boumsell L. Biochemical analysis and epitope mapping of mAb defining CD39. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:383-385.
  5. Häusler SF, Del Barrio IM, Diessner J, et al. Anti-CD39 and anti-CD73 antibodies A1 and 7G2 improve targeted therapy in ovarian cancer by blocking adenosine-dependent immune evasion.. Am J Transl Res. 2014; 6(2):129-39. (Clone-specific: Flow cytometry, Functional assay, Inhibition). View Reference
  6. Jones M, Mason DY. CD39 Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:157-159.
View All (6) View Less
567534 Rev. 1

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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.