Standard flow cytometry methods were used to evaluate cell surface marker expression using various combinations of FITC, PE, PE-Cy5, PE-Cy7, APC, APC-Cy7–conjugated anti-CD3, anti-CD4, anti-CD16, anti-CD25, anti-CD29, anti-CD45RA, anti-CD56, and PE anti-CXCR4 antibodies (Life Technologies; the rest of the antibodies are from BD). CFSE dilution assays were performed as previously described (Zhang et al., 2009 (link)). Samples were acquired on a FACSCanto instrument using FACSDiva software (BD). Analyses were performed using the FlowJo software version 9 and higher (Tree Star).
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Comprehensive Immune Phenotyping by Flow Cytometry
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Corresponding Organization : ARC Centre of Excellence in Advanced Molecular Imaging
Other organizations : MRC Human Immunology Unit, University of Oxford
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Variable analysis
independent variables
- Various combinations of FITC, PE, PE-Cy5, PE-Cy7, APC, APC-Cy7–conjugated anti-CD3, anti-CD4, anti-CD16, anti-CD25, anti-CD29, anti-CD45RA, anti-CD56, and PE anti-CXCR4 antibodies
- Cell surface marker expression
- CFSE dilution assays were performed as previously described (Zhang et al., 2009)
- No positive or negative controls were explicitly mentioned in the provided information.
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