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3 protocols using ccr7 cd197 pe cy7

1

Phenotypic Characterization of Regulatory T Cells

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The following anti-human monoclonal antibodies (mAbs) were used for flow cytometry: CD4 Alexa Fluor® 700, CD8 APC, CD39 PE-Cy7, CD73 FITC, and CD73 eFluor450, CCR7 (CD197) PE-Cy7, PD1 (CD279) PE (eBioscience); CCR7 PE-CF594, CD45 AmCyan and CD45 FITC (Becton Dickinson), CD3 APC-H7, CD25 FITC (BioLegend) and CD25 PE (MACS Miltenyi). All mAbs were titrated using normal PBL to establish optimal dilution. Treg were defined by their ability to produce extracellular adenosine as previously described by Deaglio and Borselino [28 (link), 29 (link)]. We have previously demonstrated that these CD4+ CD39+ CD25+ Treg are FOXP3+ CD127neg [24 (link)].
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2

Multiparametric Flow Cytometry of Immune Cells

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The following anti-human monoclonal antibodies (mAb) were used for flow cytometry: CD4 AF700 (Cat 56-0048-82), CD73 FITC (Cat 11-0739-42), CD73 PE (Cat 12-0739-42), CD39 PE-Cy7 (Cat 25-0399-42), PD1 (CD279) PE (Cat 12-2799-42), and CCR7 (CD197) PE-Cy7 (Cat 25-1979-42) mAb (all eBioscience, San Diego, CA); CD5 AF700 (Cat 561159), CD19 PE-Cy7 (Cat 555414), and IgM PE (Cat 555783) (all BD Pharmingen, Heidelberg, Germany); CD26 APC (Cat 302710), CD25 FITC (Cat 356105), and CD40 PE-Cy7 (Cat 334322) (all Biolegend, San Diego, CA); CD25 PE (Cat 130-101-426) and IgA APC (Cat 130-093-113) (Milenty Biotec, Bergisch Gladbach, Germany). All mAb were titrated using normal PBMC to establish optimal staining dilutions.
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3

Multiparametric Flow Cytometry Panel

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The following anti-human monoclonal antibodies (mAbs) were used for flow cytometry: CD4 Alexa Fluor ® 700, CD8 APC, CD39 PE-Cy7, CD73 FITC, and CD73 eFluor450, CCR7 (CD197) PE-Cy7, PD-1 (CD279) PE (eBioscience); CD19 PE-Cy5, CCR7 PE-CF594, CD45 AmCyan and CD45 FITC (Becton Dickinson), CD3 APC-H7, CD25 FITC, CD25 PE (MACS Miltenyi). All mAbs were titrated using PBMC of healthy donors to establish optimal dilution.
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