After 5 days of culture, splenocytes were stained with fluorescein isothiocyanate-conjugated anti-CD4 (eBioscience, Fisher Scientific, Loughborough, UK) and Pacific blue-conjugated anti-CD25 (Biolegend, London, UK) antibodies, permeabilized and stained with phycoerythrin-conjugated anti-FoxP3 (eBioscience). Samples were analyzed on an LSR Fortessa™ flow cytometer (BD Bioscience, Oxford, UK) using FACSDiva™ software. Regulatory T cells were defined as CD4+CD25+FoxP3+.40 (link)
Phycoerythrin conjugated anti foxp3
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom, United States
Phycoerythrin-conjugated anti-FoxP3 is a fluorescent-labeled antibody reagent for the detection and analysis of FoxP3-expressing cells by flow cytometry. FoxP3 is a transcription factor that serves as a marker for regulatory T cells. The phycoerythrin (PE) fluorescent label allows for the identification and quantification of FoxP3-positive cells in a sample.
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2 protocols using phycoerythrin conjugated anti foxp3
1
Regulatory T Cell Identification
2
Identification of Th17 and Treg cells in AS
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Spleen tissues were obtained from SKG mice in which AS had been induced by curdlan injection at 6 weeks of age. To identify populations of Th17 and Treg cells, the tissues were reacted with fluorescein isothiocyanate-conjugated anti-CD4, allophycocyanin-conjugated anti-IL-17, allophycocyanin-conjugated anti-CD25, phycoerythrin-conjugated anti-FOXP3, and Alexa Fluor 405-conjugated anti-CCR9 antibodies (eBioscience, San Diego, CA, USA). Stained tissue sections were observed under a confocal microscope (LSM 510Meta; Carl Zeiss, Oberkochen, Germany). Stained cells were enumerated using Pannoramic MIDI and Pannoramic viewer (3D Histech Ltd., Hungary).
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