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OMIP-020

Phenotypic characterization of human γδ T-cells by multicolor flow cytometry

Kilian Wistuba-Hamprecht, Graham Pawelec, Evelyna Derhovanessian Cytometry PART A, Volume 85, Issue 6, 522-524 (2013)

PURPOSE: This panel was composed and optimized to investigate the differentiation stages of human γδ T-cells in cryopreserved peripheral blood mononuclear cells (PBMC) from healthy individuals (Tables 1 and 2). As the majority of pan-γδ T-cell antibodies available commercially proved to be inappropriate for detecting all γδ T-cell populations in combination with other markers, this panel provides an essential tool for the analysis of different subsets of human γδ T-cells by flow cytometry. The panel works very well with cryopreserved PBMC. Other tissues have not been tested.

CELL TYPE: PBMC

MACHINE: BD LSR2

Marker Target Clone Fluorochrome Purpose
Dead cells Human EMA viability
CD3 Human UCHT1 Alexa Fluor 700 Lineage
CD4 Human OKT4 PE-Cy7 CD4 T cell
CD8 Human SK1 APC-H7 CD8 T cell
CD16 Human 3G8 Brilliant Violet 711 ?dT activation
CD27 Human APC differentiation
CD28 Human CD28.2 PE differentiation
CD45RA Human Pacific Blue differentiation
TCR gamma/delta Human 11F2 Purified ?dT cell
IgG Human N/A Pacific Orange secondary reagent to ?d-TCR (clone qqF2)
TCR V delta 1 Human TS8.2 FITC ?dT cell subset
TCR V delta 2 Human B6 PerCP ?dT cell subset