< OMIPS Directory
OMIP-015

Human regulatory and activated T-cells without intracellular staining

Yolanda D. Mahnke, Margaret H. Beddall, Mario Roederer Cytometry PART A, Volume 83A, Issue 2, 179-181 (2013)

PURPOSE: The present panel was optimized to investigate the frequency and phenotype of regulatory T-cells (Treg), as well as the activation status of CD4+ and CD8+ T-cells in peripheral blood mononuclear cells (PBMC) from healthy individuals, without the use of intracellular staining (i.e., excluding the use of the canonical Treg marker, FoxP3). The panel has been developed using cryopreserved PBMC and we have observed similar results with fresh specimens. Other tissue types have not been tested.

CELL TYPE: PBMC

MACHINE: BD LSR2

Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Aqua viability
CD25 Human M-A251 PE-Cy5 Treg
CD38 Human HIT2 PE-CF594 activation/differentiation
CD39 Human PE-Cy7 Treg function
CD45RO Human UCHL1 FITC activation/differentiation
CD73 Human AD2 PE Treg function
CD127 Human eBioRDR5 APC-eFluor 780 Treg
PD-1 Human EH12.2H7 Brilliant Violet 421 activation/differentiation
CD4 Human OKT4 Qdot 605 lineage
CD8 Human RPA-T8 Qdot 585 lineage
CD45RA Human 5H9 Qdot 705 activation/differentiation
HLA-DR Human G46-6 Alexa Fluor 680 activation/differentiation