Anti mouse cd86 fitc

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).

BMDCs of each treatment group in experiment (1) were collected and were divided into two tubes, to which anti-mouse MHCII-PE-(yanin5-conjugated) Cy5, anti-mouse CD80-FITC, and anti-mouse CD11c-PE (phycoerythrin), anti-mouse CD86-FITC, and anti-CD40-PE-cy5 (eBioscience), respectively, were added, followed by the addition of 500 μl of 2% PFA in PBS, pH 7·4. A blank control was included. All samples were incubated in the dark at 4°C for 30 min, and were then subjected to flow-cytometry detection of surface molecules of BMDCs. Data were collected for at least 1×10⁴ cells, and the experiment was carried out four times.