OMIP-006
Phenotypic subset analysis of human T regulatory cells via polychromatic flow cytometry
Cytometry PART A, Volume 81A, Issue 4, 281-283 (2012)
David M. Murdoch, Janet S. Staats, Kent J. Weinhold
Description: This panel was optimized for the enumeration and phenotypic characterization of T regulatory cells (Tregs) within the CD4+ T-cell pool using human peripheral blood mononuclear cells (PBMC) using intranuclear and intracellular staining methods. The panel was optimized for HIV+ clinical trial specimens through the use of HIV-infected and normal donor PBMC. Because the panel is to be used in the context of testing cryopreserved PBMC obtained from multiple sites participating in clinical trials, it was essential to develop an assay that performed well using cryopreserved PBMC. Other tissue types have not been tested.
Species: Human
Tissue: Peripheral blood
Cell Subset: T cell, Treg
Keyword: Cryopreserved pbmc, Hiv
Machine: LSR II
Panel details
| Marker | Species | Clone | Fluorophore | Purpose | Co-expressions groups | Antigen Density |
|---|---|---|---|---|---|---|
| Dead cells | Human | Live/Dead Fix Violet | viability | |||
| CD4 | Human | SK3 | PerCP-Cy5.5 | T cell subset | T reg | High |
| FOXP3 | Human | PCH101 | PE | Treg identification | T reg | Low |
| CD25 | Human | B1.49.9 | ECD | T cell activation, Treg identification | T reg | Low |
| CD49d | Human | 9F10 | PE-Cy5 | Treg subset | T reg | Low |
| Helios | Human | 22F6 | Alexa Fluor 647 | Treg subset | T reg | Medium |
| CD3 | Human | UCHT1 | V500 | T cell subset | T reg | High |
| CD45RO | Human | UCHL1 | FITC | maturation/differentiation | T reg | Medium |
| CD39 | Human | eBioA1 (A1) | PE-Cy7 | Treg subset | T reg | Low |
