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BV421 Rat Anti-Mouse CD39
BV421 Rat Anti-Mouse CD39

Flow cytometric analysis of CD39 expression on mouse splenic lymphocytes. Balb/c mouse splenocytes were stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553729, 557307, or 561828), APC Rat Anti-Mouse CD25 (Cat. No. 557192 or 561048), and BD Horizon™ BV421 Rat IgG2b, κ Isotype Control (Cat. No. 562603) or BD Horizon™ BV421 Rat Anti-Mouse CD39 antibody (Cat. No. 567105) at 0.5 μg/test. 7-AAD (7-Amino-Actinomycin D) Solution (Cat. No. 559925) was added to cells right before analysis. Upper panels: Two-parameter flow cytometric pseudocolor plots showing correlated expression of CD39 (right) or Ig Isotype control (left) versus CD4 staining were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphocytes. Lower panels: Two-parameter flow cytometric pseudocolor plots showing correlated expression of CD39 (right) or Ig Isotype control (left) versus CD25 staining were derived from CD4-positive viable lymphocytes. Flow cytometry and data analysis were performed using a BD X-20 LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Flow cytometric analysis of CD39 expression on mouse splenic lymphocytes. Balb/c mouse splenocytes were stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553729, 557307, or 561828), APC Rat Anti-Mouse CD25 (Cat. No. 557192 or 561048), and BD Horizon™ BV421 Rat IgG2b, κ Isotype Control (Cat. No. 562603) or BD Horizon™ BV421 Rat Anti-Mouse CD39 antibody (Cat. No. 567105) at 0.5 μg/test. 7-AAD (7-Amino-Actinomycin D) Solution (Cat. No. 559925) was added to cells right before analysis. Upper panels: Two-parameter flow cytometric pseudocolor plots showing correlated expression of CD39 (right) or Ig Isotype control (left) versus CD4 staining were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphocytes. Lower panels: Two-parameter flow cytometric pseudocolor plots showing correlated expression of CD39 (right) or Ig Isotype control (left) versus CD25 staining were derived from CD4-positive viable lymphocytes. Flow cytometry and data analysis were performed using a BD X-20 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 Horizon™
ectonucleoside triphosphate diphosphohydrolase 1; Entpd1; ecto-ATP diphosphohydrolase 1; ecto-ATPDase 1; ecto-ATPase 1; Cd39; NTPDase 1
Mouse (QC Testing)
Rat F344, also known as Fischer, CDF IgG2b, κ
Mouse CD39 Transfected Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2870022
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 to the dye under optimum conditions and unconjugated antibody and free dye 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 cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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. 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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  9. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567105 Rev. 2
Antibody Details
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Y23-1185

The Y23-1185 monoclonal antibody specifically recognizes mouse CD39, also known as ectonucleoside triphosphate diphosphohydrolase 1 (NTPDase 1), which is an enzyme on the surface of vascular endothelial cells, antigen presenting cells and activated immune cells. CD39 is encoded by ENTPD1 which belongs to the ectoenzyme family. The catalytic portion of the CD39 is extracellular, where it 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.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon™ BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon™ BV421, Pacific Blue, and BD Horizon™ V450 cannot be used simultaneously.

567105 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
567105 Rev.2
Citations & References
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Development References (5)

  1. Allard D, Allard B, Stagg J. On the mechanism of anti-CD39 immune checkpoint therapy. J Immunother Cancer. 2020; 8(1):e000186. (Biology). View Reference
  2. 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
  3. Mizumoto N, Kumamoto T, Robson SC, et al. CD39 is the dominant Langerhans cell-associated ecto-NTPDase: modulatory roles in inflammation and immune responsiveness.. Nat Med. 2002; 8(4):358-365. (Biology). View Reference
  4. Salmi M, Jalkanen S. Ectoenzymes controlling leukocyte traffic.. Eur J Immunol. 2012; 42(2):284-92. (Biology). View Reference
  5. Zhou Q, Yan J, Putheti P, et al. 2009; 9(10):2303-2311. (Biology). View Reference
View All (5) View Less
567105 Rev. 2

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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.