Multicolor Flow Cytometric Analysis of Tregs

Multicolor flow cytometric analysis of Tregs was performed as described previously [17 (link)]. Monoclonal antibodies (mAbs) with specificity against CD4 (RM4-4), CD25 (7D4), CD62L (Mel-14), and CTLA4 (UC10-4F10-11) were used. For intracellular staining, FoxP3 (FJK-16s) Staining Kit (eBioscience) was used according to the manufacturer's protocol. PI was used for dead cell exclusion when appropriate. Surface staining was performed according to standard procedures, and flow cytometric analysis was done on Coulter Cytomics FC500 using CXP software (Coulter, Austria) for acquisition and analysis.

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Pilat N., Mahr B., Gattringer M., Baranyi U, & Wekerle T. (2018). CTLA4Ig Improves Murine iTreg Induction via TGFβ and Suppressor Function In Vitro. Journal of Immunology Research, 2018, 2484825.

Publication 2018

FoxP3 (FJK-16s) Staining Kit Cytomics FC500

Antibodies specificity Cd25 Cd62l Cell Ctla4 Flow cytometric Intracellular

Corresponding Organization : Medical University of Vienna

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Variable analysis

independent variables

Monoclonal antibodies (mAbs) with specificity against CD4 (RM4-4), CD25 (7D4), CD62L (Mel-14), and CTLA4 (UC10-4F10-11)

dependent variables

Multicolor flow cytometric analysis of Tregs

control variables

Surface staining performed according to standard procedures
Flow cytometric analysis done on Coulter Cytomics FC500 using CXP software (Coulter, Austria) for acquisition and analysis

controls

PI was used for dead cell exclusion when appropriate

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