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Miltenyi Biotec distribution:
As a global market leader with numerous subsidiaries and distributors, Miltenyi Biotec is committed to providing our customers around the world with the highest quality products. In addition to direct selling in more than 20 countries in North America, Europe and Asia/Pacific, Miltenyi Biotec also provides support for our customers through an extensive distributor network covering dozens of additional countries.
Human peripheral blood mononuclear cells (PBMCs) were isolated with the CD4+
T Cell Isolation Kit, followed by a CD25 /CD45RO depletion. Afterwards, cells were either left unstimulated (left images) or stimulated with CD3 and CD28 antibodies as well as with human IL-4 and human IL-2 for three days. Cells were then polarized with human IL-4 and human IL-2 and restimulated with PMA and ionomycin. After two hours brefeldin A was added.
Cells were fixed, permeabilized, and intracellularly stained with Anti-IL-9 antibodies as well as with CD4 and CD154 antibodies. CD4+
cells were pre-gated for the analysis. Cells were then analyzed by flow cytometry using the MACSQuant®
Analyzer. Cell debris were excluded from the analysis based on scatter signals.