T cells were first stained with LIVE/DEAD® Fixable Aqua Dead Cell Stain Kit (Thermo Fisher, UK) before being stained with conjugated antibodies, with each step incubation for 20 min at 4°C. Antibodies used for surface staining used included the following: PerCP/Cy5.5-CD8 (SK1; BD Biosciences; RRID:AB_2687497 ), PE/Cy7-CD27 (M-T271; BioLegend; RRID:AB_2562258 ), BV421-CCR7 (G043H7; BioLegend; RRID:AB_11203894 ), BV711-CD45RA (HI100; BioLegend; RRID:AB_11218999 ), and fluorescein isothiocyanate (FITC)-CD28 (CD28.2; BD Biosciences; RRID:AB_396071 ). All samples were acquired on BD LSR Fortessa (BD Biosciences) flow cytometer and analyzed using FlowJo™ v.10 software (Tree Star Inc.).
Percp cy5.5 cd8 sk1
Manufactured by BD
PerCP/Cy5.5-CD8 (SK1) is a fluorescently labeled antibody that binds to the CD8 protein expressed on the surface of T lymphocytes. It is used in flow cytometry applications to identify and quantify CD8+ T cells within a sample.
Automatically generated - may contain errors
2 protocols using percp cy5.5 cd8 sk1
1
Multiparametric T Cell Immunophenotyping
2
T cell Cytokine Profiling Protocol
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T cells were treated with Monensin and Brefeldin A (BD Biosciences) prior to co-culture with peptide-stimulated lymphoblastoid B cell line (BCL) or HCT116 for 5 h at 37°C. Cells were then surface stained with PerCP/Cy5.5-CD8 (SK1; BD Biosciences; RRID:AB_2687497 ) before being fixed with Cytofix/Cytoperm™ (BD Biosciences) and stained with conjugated intracellular antibodies including Alexa Flour488-IFNγ (B27; BD Biosciences; RRID:AB_396827 ) and APC-TNFα (Mab-11; BioLegend; RRID:AB_315264 ). Samples were acquired on an Attune Nxt flow cytometer (Thermo Fisher Inc.) and analyzed using FlowJo™ v.10 software (Tree Star Inc.).
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