| Assay Method Information | |
| | Fluorescence Cell-Based Secondary Assay to Measure Toxicity of Compounds Not in the Presence of Fluconazole |
| Description: | 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 NIH 3T3 mammalian fibroblasts cultured in 384-well format in the presence of a sub-toxic concentration of Fluconazole. Test compounds that do not inhibit subsequent growth in the presence of Fluconazole will merit further evaluation for their non-toxicity. This whole cell phenotypic screening approach will only capture compounds that are capable of entering and accumulating in cells to bioactive concentrations. Compound activity will be measured by the metabolism of Alamar Blue, a cell stain that is metabolized to a fluorescent product by living cells but that remains non-fluorescent in wells with growth-inhibited organisms. Probe attributes: a. Compounds that inhibit yeast growth in the presence, but not in the absence of 8 ug/ ml Fl |
| Affinity data for this assay | |
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