Multiparameter flow cytometric analysis of CD39 (ENTPD1) expression on Human peripheral blood leukocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with either BD Horizon™ RB744 Mouse IgG2b, κ Isotype Control (Cat. No. 570523; Left Plot) or BD Horizon™ RB744 Mouse Anti-Human CD39 (ENTPD1) antibody (Cat. No. 570637/570723; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD39 (ENTPD1) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ software.
Multicolor flow cytometric analysis of CD39 (ENTPD1) expression on Human peripheral blood lymphocytes. Human peripheral blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD4 (Cat. No. 562424), Alexa Fluor™ 700 Mouse Anti-Human CD25 (Cat. No. 561398), and PE Anti-Human CD127 (Cat. No. 557938) antibodies, and with either BD Horizon™ RB744 Mouse IgG2b, κ Isotype Control (dashed line histogram; Right Plot) or BD Horizon™ RB744 Mouse Anti-Human CD39 (ENTPD1) antibody (solid line histogram; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histogram showing CD39 (ENTPD1) expression (or Ig Isotype control staining) [Right Plot] was derived from CD4+CD25+CD127low gated events (ie, cells with a Regulatory T cell immunophenotype; Left Plot) with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
Multiparameter flow cytometric analysis of CD39 (ENTPD1) expression on Human peripheral blood leukocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with either BD Horizon™ RB744 Mouse IgG2b, κ Isotype Control (Cat. No. 570523; Left Plot) or BD Horizon™ RB744 Mouse Anti-Human CD39 (ENTPD1) antibody (Cat. No. 570637/570723; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD39 (ENTPD1) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ software.
Multicolor flow cytometric analysis of CD39 (ENTPD1) expression on Human peripheral blood lymphocytes. Human peripheral blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD4 (Cat. No. 562424), Alexa Fluor™ 700 Mouse Anti-Human CD25 (Cat. No. 561398), and PE Anti-Human CD127 (Cat. No. 557938) antibodies, and with either BD Horizon™ RB744 Mouse IgG2b, κ Isotype Control (dashed line histogram; Right Plot) or BD Horizon™ RB744 Mouse Anti-Human CD39 (ENTPD1) antibody (solid line histogram; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histogram showing CD39 (ENTPD1) expression (or Ig Isotype control staining) [Right Plot] was derived from CD4+CD25+CD127low gated events (ie, cells with a Regulatory T cell immunophenotype; Left Plot) with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
Product Details
BD Horizon™
ENTPD1; NTPDase-1; Ecto-ATPase 1; Ecto-ATPDase 1
Human (QC Testing)
Mouse IgG2b, κ
Flow cytometry (Routinely Tested)
5 µl/test
IV A54
953
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
RB744 Mouse IgG2b, κ Isotype Control RUO
Size
0.1 mg
Cat No.
570523
BV421 Mouse Anti-Human CD4 RUO
Size
100 Tests
Cat No.
562424
Alexa Fluor® 700 Mouse Anti-Human CD25 RUO
Size
50 Tests
Cat No.
561398
PE Mouse Anti-Human CD127 RUO
Size
0.1 mg
Cat No.
557938
570723 Rev. 1
Antibody Details
TU66
The TU66 monoclonal antibody specifically recognizes human CD39 which is also known as Ectonucleoside triphosphate diphosphohydrolase 1 (NTPDase 1), Ecto-ATP diphosphohydrolase 1 (Ecto-ATPDase 1), or Ecto-apyrase. CD39 is an integral membrane glycoprotein with two transmembrane domains, N- and C-terminal cytoplasmic tails, and an extracellular region that contains the NTPDase 1 active site. CD39 is encoded by ENTPD1 which belongs to the ectoenzyme family. CD39 is variably expressed on activated T cells and B cells, regulatory T cells (Treg), dendritic cells, Langerhans cells, NK cells, monocytes, macrophages, endothelial cells, and granulocytes. CD39 acts on extracellular nucleoside triphosphates and diphosphates including ATP and ADP that are hydrolyzed into AMP. Through cell surface CD73 (Ecto-5'-nucleotidase), regulatory T cells can act on extracellular AMP to generate immunosuppressive adenosine. CD39 is involved in the control of the extracellular pool of phosphorylated nucleosides, the suppression of inflammation and immunity, and the regulation of platelet activation.
570723 Rev. 1
Format Details
RB744
The BD Horizon RealBlue™ 744 (RB744) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 746-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB744 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), we recommend using an optical filter centered near 750-nm (e.g., a 750/60-nm bandpass filter).
RB744
Blue 488 nm
498 nm
746 nm
570723 Rev.1
Citations & References
View product citations for antibody "570723" on CiteAb
Development References (5)
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Borsellino G, Kleinewietfeld M, Di Mitri D, et al. Expression of ectonucleotidase CD39 by Foxp3+ Treg cells: hydrolysis of extracellular ATP and immune suppression.. Blood. 2007. (Biology). View Reference
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Duensing S, Kirshner H, Atzpodien J. CD39 as a novel marker of in vivo immune activation. Blood. 1994; 83(12):3826-3827. (Biology). View Reference
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Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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Stein H, Lennert K, Mason DY, Liangru S, Ziegler A. Immature sinus histiocytes. Their identification as a novel B-cell population. Am J Pathol. 1984; 117(1):44-52. (Clone-specific: Immunohistochemistry). View Reference
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Ziegler A, Uchanska-Ziegler B, Stein H, Hadam M. A mAb A54 (Tü 66) recognizing a novel avtivation antigen. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:467-468.
570723 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.