Cd39 percp efluor710

Manufactured by Thermo Fisher Scientific

The CD39 PerCP-eFluor710 is a fluorescent-labeled antibody used for the detection and identification of CD39-expressing cells in flow cytometry analysis. CD39 is an enzyme involved in the regulation of extracellular ATP and adenosine levels. This product provides a tool for researchers to study the role of CD39 in various biological processes.

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2 protocols using cd39 percp efluor710

Comprehensive Flow Cytometry Analysis

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The following antibodies were used for flow cytometry analysis: CD11c APC, CD80 FITC, CD83 FITC, CD86 PE, HLA-DR FITC, CD1a FITC, CD14 FITC, CD4PE, CD4 PE-Cy7, CD45RA PE, CD45RO FITC, CD25 PE-Cy7, CTLA-4 APC, CD39 PerCP-eFluor710, FOXP3 Alexa Fluor 700, IL-10 PE, IFN-γ PE-Cy7, and IL-17 APC (all eBioscience). For surface staining, cells were incubated in PBS 10% FBS containing the respective antibodies for 30 min at 4°C, washed, and stored in IC fixation buffer until analysis (eBioscience). Intracellular FoxP3 was detected using FoxP3-staining kit (eBioscience) according to the manufacturer’s instructions. For intracellular cytokines detection, cells were treated with 50 ng/ml PMA, 1 μg/ml ionomycin, and 1 μl/ml brefeldin A for 5 h. After harvesting, cells were surface stained for CD4. Intracellular cytokine staining was performed in permeabilization buffer (eBioscience) before cells were washed and resuspended in FACS buffer for flow cytometry analysis. Cell viability was assessed by 7-AAD and annexin-V PE staining (eBioscience). Data were collected on FACSCalibur and FACSAria cytometers (Beckton Dickinson, San Diego, CA, USA) and analyzed with Weasel v3.0.2 software.

Maggi J., Schinnerling K., Pesce B., Hilkens C.M., Catalán D, & Aguillón J.C. (2016). Dexamethasone and Monophosphoryl Lipid A-Modulated Dendritic Cells Promote Antigen-Specific Tolerogenic Properties on Naive and Memory CD4+ T Cells. Frontiers in Immunology, 7, 359.

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Comprehensive Flow Cytometry Analysis of Th1 Cells

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Th1 cells were washed with PBS supplemented with 0.1% BSA and 0.01% sodium azide, and stained using CD4 APC (clone: RPA-T4), CD39 PerCP-eFluor 710 or FITC (clone: eBio(A1) from eBioscience), and CD45 PE/FITC/PE-Cy7 (clone: 2D1; H130). For assessment of cell death, Th1 cells were stained for surface markers (CD4 PE and CD39 FITC), re-suspended in Annexin V buffer, and stained with Annexin V APC and 7AAD according to manufacturer's instructions. For in vivo monitoring of human Th1 cells in the spleen, cells were stained with human CD45 PE-Cy7, CD4 Pacific Blue, and CD39 PerCP-eFluor710. For intra-cellular (IC) flow cytometry, fixation and permeabilization buffer was utilized (eBioscience). IC flow cytometry was performed with combinations of CD45 FITC, CD4 Pacific Blue, CD39 PerCP-eFluor 710, and FoxP3 PE (clone 249D). For IC cytokine flow cytometry, human Th1 cells from spleen, bone marrow, lung, liver, and skin were stimulated with PMA (50ng/ml) and ionomycin (0.5mg/ml) for 4hrs together with Golgiplug and Golgistop (BD Biosciences) for the final 2hrs. Cells were subsequently fixed and stained with CD45 PE-Cy7, CD4 Pacific Blue, CD39 PerCP-eFluor 710, IL-2 FITC (clone: MQ1-17H12), IFN-γ PE (clone: 45.B3), TNF-α APC (clone: MAB11; eBioscience). Cells after staining were examined on an LSRII (Becton Dickinson) and data analyzed using FlowJo 9.7.6 (Treestar).

Amarnath S., Foley J.E., Farthing D.E., Gress R.E., Laurence A., Eckhaus M.A., Métais J.Y., Rose J.J., Hakim F.T., Felizardo T.C., Cheng A.V., Robey P.G., Stroncek D.E., Sabatino M., Battiwalla M., Ito S., Fowler D.H, & Barrett A.J. (2015). Bone Marrow Derived Mesenchymal Stromal Cells Harness Purinergenic Signaling to Tolerize Human Th1 Cells In Vivo. Stem cells (Dayton, Ohio), 33(4), 1200-1212.

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