Draining lymph node cells were isolated from the axillary lymph nodes at the end of the experiments and stimulated ex vivo with 50 ng ml−1 PMA and 1 μg ml−1 ionomycin in the presence of Monensin for 5 h, followed by intracellular staining for IL-13-PE (eBiosciences, clone Bio13A) or IL-17A-APC/Cy7 (Biolegend, clone TC11-18H10.1). The frequency of cytokine producing CD4+ T cells was determined by flow cytometry.
Il 17a apc cy7
Manufactured by BioLegend
IL-17A-APC/Cy7 is a fluorochrome-conjugated antibody that binds to the interleukin-17A (IL-17A) protein. IL-17A is a pro-inflammatory cytokine produced by T helper 17 (Th17) cells and other immune cells. The APC/Cy7 fluorophore combination provides a bright signal for flow cytometric detection and analysis.
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Lab products found in correlation
Permeabilization buffer, by Thermo Fisher Scientific (1 mentions)
Il 4 af488, by Thermo Fisher Scientific (1 mentions)
Brefeldin a, by Thermo Fisher Scientific (1 mentions)
Il 10 bv421, by BioLegend (1 mentions)
Tnf α percp cy5, by BioLegend (1 mentions)
Fixation permeabilization buffer, by Thermo Fisher Scientific (1 mentions)
2 protocols using il 17a apc cy7
1
Characterizing Cytokine-Producing CD4+ T Cells
2
Phenotypic Analysis of Activated T Cells
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To assess their functional phenotype, 106 T cells were activated with 50 ng/mL phorbol myristic acid and 1 µg/mL ionomycin for 30 min followed by incubation for 3 h at 37°C in the presence brefeldin A (eBioscience). Cells were fixed in fixation/permeabilization buffer (eBioscience), washed in permeabilization buffer (eBioscience), stained using anticytokine antibodies (IL-4 AF488 from eBioscience and interferon-γ AF700, IL-10 BV421, IL-17A APC/Cy7, and tumor necrosis factor (TNF)-α PerCP-Cy5.5 from BioLegend) and analyzed on an LSR II (BD Biosciences) and using FlowJo software (TreeStar Inc.).
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