Thymus Cell Suspension Preparation and Staining

Thymus cell suspensions were prepared and stained as described (Hager-Theodorides et al., 2005 (link)) using combinations of the following directly conjugated antibodies at concentration of 1:100: (from BD Pharmingen) anti-γδPE (catalogue no. 553178); from eBioscience: anti-TCRβFITC (catalogue no. 11-5961-85), antiCD3PE (catalogue no. 12-0031-82), anti-CD24PE (catalogue no. 12-0241-82) and anti-CD69FITC (catalogue no. 11-0691-85); (from Biolegend) anti-CD3FITC (catalogue no. 100204), anti-CD5FITC (catalogue no. 100605), anti-Qa2FITC (catalogue no. 121709), anti-CD4APC (catalogue no. 116014), anti-CD5PE (catalogue no. 100607), anti-CD8PerCP/Cy5.5 (catalogue no. 100734), anti-CD4PerCP/Cy5.5 (catalogue no. 100539) and anti-CD8APC (catalogue no. 100712). Data were acquired on a C6 Accuri flow cytometer (BD Biosciences) and analysed using FlowJo software. Live cells were gated by FSC and SSC profiles. Data represent at least three experiments.

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Solanki A., Yanez D.C., Ross S., Lau C.I., Papaioannou E., Li J., Saldaña J.I, & Crompton T. (2018). Gli3 in fetal thymic epithelial cells promotes thymocyte positive selection and differentiation by repression of Shh. Development (Cambridge, England), 145(3), dev146910.

Publication 2018

anti-TCRβFITC antiCD3PE anti-CD24PE anti-CD69FITC anti-CD3FITC anti-CD5FITC anti-CD4APC anti-CD5PE anti-CD8PerCP/Cy5.5 anti-CD4PerCP/Cy5.5 anti-CD8APC C6 Accuri flow cytometer

Antibodies Cells Cy5 5 Thymus

Corresponding Organization :

Other organizations : University College London, University of East London

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Variable analysis

independent variables

Staining with combinations of directly conjugated antibodies

dependent variables

Thymus cell populations measured by flow cytometry

control variables

Preparation and staining of thymus cell suspensions as described in Hager-Theodorides et al., 2005
Antibody concentrations at 1:100
Live cells gated by FSC and SSC profiles

controls

No positive or negative controls were explicitly mentioned in the provided information.

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