Multicolor Flow Cytometry Immunophenotyping

Cell staining was performed as previously described^{34 (link)}. Cells were washed in cold PBS with 2% FCS and 0.1% sodium azide and following stained with fluorescein isothiocyanate (FITC)-, phycoerythrin (PE)-conjugated anti-human monoclonal antibodies which were including anti-CD3/CD16 + CD56 (APC/FITC), NKp46, CD69, CD62L, CD158b, and CD158e (PE) from Becton-Dickinson for flow cytometric analysis. The fluorescent staining was analyzed on a Canto II (BD Biosciences) flow cytometer. Electronic gates were set to enable analysis of the fluorescence of the viable cell population according to FSC/SSC histograms following anti-CD3/ CD16 + CD56 stimulation. The percentage of cells stained with each monoclonal antibody was determined by comparing each histogram with one from control cells stained with FITC- or PE- labeled isotype control monoclonal antibodies.

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Lin S.J., Hsu C.Y., Kuo M.L., Lee P.T., Hsiao H.S, & Chen J.Y. (2020). Phenotypic and functional characterization of natural killer cells in rheumatoid arthritis-regulation with interleukin-15. Scientific Reports, 10, 5858.

Publication 2020

CD62L Canto II

Anti cd3 Cd158e Cd62l Cells Cold Flow cytometric Fluorescence Human Isothiocyanate Isotype Monoclonal antibodies Nkp46 Phycoerythrin Sodium azide

Corresponding Organization : Chang Gung Memorial Hospital

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Variable analysis

independent variables

Cell staining protocol

dependent variables

Percentage of cells stained with each monoclonal antibody

control variables

Cells were washed in cold PBS with 2% FCS and 0.1% sodium azide
Analysis of the fluorescence of the viable cell population according to FSC/SSC histograms following anti-CD3/ CD16 + CD56 stimulation
Comparison of each histogram with one from control cells stained with FITC- or PE- labeled isotype control monoclonal antibodies

positive controls

Cells stained with anti-CD3/CD16 + CD56 (APC/FITC) monoclonal antibody

negative controls

Cells stained with FITC- or PE- labeled isotype control monoclonal antibodies

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