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T cells were isolated from human peripheral blood mononuclear cells (PBMCs). Cells were then stimulated with 50 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 6 hours and then positively selected for their IL-17A expression. After cultivation for 11 days, cells were restimulated with 50 ng/mL PMA and 1 µg/ml Ionomycin for 6 hours, followed by an incubation with 1 µg/ml Brefeldin A for 4 hours. Cells were then fixed, permeabilized and stained with the REA Control (I) antibodes (left image) or with Anti-IL-17F antibodies. Flow cytometry was performed with the MACSQuant®
Analyzer. Cell debris were excluded from the analysis based on scatter signals.
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