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Protocol Steps  >  adding substance to cell culture step  >  Phagocytosis assay

Phagocytosis assay

adding substance to cell culture step

Protocol: Internalization assay

Yeasts/spores are biotinylated using 10 mg/ml sulfo-NHS-LC-biotin (Sigma-Aldrich) in 50 mM NaHCO3 pH 8.5 for 2 hours at 4°C. The remaining reactive biotin molecules are inactivated by incubation in 100 mM Tris-HCl pH 8.0 for 40 minutes at 4°C. DCs are then treated with biotinylated spores/yeasts. After 1 hour, cells are permeabilized and labeled with aHLA-DR-FITC. After zymolyase treatment, intracellular yeasts/spores are detected using APC-labeled streptavidin and analyzed by flow cytometry.

created over 15 years ago (21 January 2010)    last modified over 13 years ago (28 September 2011)   [ RDF Rdf ]   [ RelFinder Relfinder ]