Comprehensive Cytometric Analysis of T Cell Subsets

Flow cytometry was performed using the following antibodies from BioLegend, unless otherwise stated. Antibodies to the following human molecules were used: CD69-AF647 (FN50), CD69-PE (FN50), CD3-BV421 (OKT3), γδTCR-PeCy7 (IMMU510; Beckman Coulter), CD45-PacificBlue (HI30), TCRVδ2-FITC (B6). Antibodies to the following murine molecules were used: TCRδ-PerCPe710 (GL3), TCRδ-APC (GL3), Vγ7-AF647 (F2.67, provided by P.Pereira). Other antibodies were as follows: DYKDDDDK-PE (FLAG, L5), HA-AF647 (16B12). 6xHis-APC (Biolegend, J095G46). Data were acquired on BD Canto II or Fortessa cytometers. sTCR staining was performed as in Melandri et al. (2018) (link).

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Willcox C.R., Vantourout P., Salim M., Zlatareva I., Melandri D., Zanardo L., George R., Kjaer S., Jeeves M., Mohammed F., Hayday A.C, & Willcox B.E. (2019). Butyrophilin-like 3 Directly Binds a Human Vγ4+ T Cell Receptor Using a Modality Distinct from Clonally-Restricted Antigen. Immunity, 51(5), 813-825.e4.

Publication 2019

CD69-PE CD45-PacificBlue BD Canto II Fortessa

Af647 Antibodies Fitc Flow cytometry Human Murine Okt3

Corresponding Organization : King's College London

Other organizations : Université de Tours

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Variable analysis

independent variables

Antibodies to the following human molecules: CD69-AF647 (FN50), CD69-PE (FN50), CD3-BV421 (OKT3), γδTCR-PeCy7 (IMMU510; Beckman Coulter), CD45-PacificBlue (HI30), TCRVδ2-FITC (B6)
Antibodies to the following murine molecules: TCRδ-PerCPe710 (GL3), TCRδ-APC (GL3), Vγ7-AF647 (F2.67, provided by P.Pereira)
Other antibodies: DYKDDDDK-PE (FLAG, L5), HA-AF647 (16B12), 6xHis-APC (Biolegend, J095G46)

dependent variables

Flow cytometry data

control variables

BD Canto II or Fortessa cytometers used for data acquisition
STCR staining performed as in Melandri et al. (2018)

positive controls

None specified

negative controls

None specified

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