| Assay Method Information | |
| | Fluorescence Cell-Based Secondary Assay of Resistant C. albicans Growth in the Presence of Fluconazole |
| Description: | Broad Institute: Reversing Antifungal Drug Resistance Project ID: 2037 Keywords: Candida albicans, drug resistance, fluconazole, Hsp90, Calcineurin, stress response Primary Collaborators: Susan Lindquist, Whitehead Institute for Biomedical Research, sll@wi.mit.edu Assay Overview: The basic assay strategy will consist of highly fluconazole-resistant C. albicans clinical isolate cultured in 384-well format in the presence of a sub-toxic concentration of fluconazole. Test compounds that inhibit subsequent growth in the presence of fluconazole will merit further evaluation for their synergy with fluconazole. This whole cell phenotypic screening approach will only capture compounds that retain activity in biological media and are capable of entering and accumulating in fungi to bioactive concentrations. Grossly cytotoxic compounds will be removed through subsequent counterscreens. Compound activity will be measured by the metabolism of Alamar Blue, a cell stain that is metabolized to |
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