Anti human ifn γ pe cy7

Anti-mouse IL-17A-PE, CD4-APC and IFN-γ-PE-cy7, anti-mouse IgG isotype, anti-human IFN-γ-PE-cy7, CD4-APC and IL-17A-PE and anti-human IgG isotype were obtained from eBioscience. For intracellular expression of IFN-γ and IL-17, CD4+ T cells were incubated for 1 hours with ionomycin (1 μg/mL), PMA (50 ng/mL) and for another 4 hours with brefeldin A (10 μg/ml, Sigma-Aldrich), harvested, washed and fixed before permeabilization.
BMDCs or MD-DCs were stained for 30 minutes at 4 °C with anti-mouse CD86-FITC (eBioscience), CD11c-APC (eBioscience), CD80-PE (eBioscience), CD40-FITC (BioLegend, San Diego, CA, USA), MHCII-PE (eBioscience), anti-human CD86-PE-cy7 (BioLegend), CD40-FITC (BioLegend), HLA-DR-PE-cy5.5 (BioLegend), and CD80-PE (BioLegend).

For IL-17 and IFN-γ staining, cultured CD4+T cells were stimulated with PMA (50ng/mL) and ionomycin (1g/mL) for 1 h at 37°C, and brefeldin A (Sigma-Aldrich) for another 4 h, then washed, fixed and permeabilized. Fluorescent anti-human CD3-APC, anti-human IL-17A-PE, anti-human IFN-γ-PE-cy7, and anti-human IgG isotype were purchased from eBiosciences.