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| Solution Information | help | |
| Enzyme: | ATP-dependent molecular chaperone HSP82 | |
| inhibitor: | BDBM68334 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Stock solutions Geldanamycin (GA) 15mM stock solution in DMSO Fluconazole (FLC, Sequoia Research Products Ltd) 2 mg/ml stock solution in PBS Pen/Strep 100x in PBS Synthetic Defined Growth Medium RPMI 1640 medium, (powder without sodium bicarbonate; Invitrogen 31800-089) Uridine 8 mg/ml in water (Sigma) Glucose 40% (w/v) in water (Sigma) MOPS Buffer (Sigma) Fungal Inoculum Test Strain: C. albicans CaCi-8 (Redding et al., 1994, Clin Infect Dis, 18: 240-242) Inoculate 500 ul of strain from cryopreserved stock into 250 ml shaker flask containing 30 ml growth medium. Shake at 30C O/N. Read OD 600 of fungal culture using standard plate reader. Dilute with Synthetic Defined Growth Medium to starting OD of the fungal inoculum of 0.00015 A600. 384-well plates (Corning). Add fluconazole stock solution (2 mg/mL in PBS) to Synthetic Defined Growth Medium to achieve 8 ug/ml. Add Pen/Strep at 0.1 ml per 10 ml media. Using Thermo Combi, dispense 20 uL/well of Synthetic Defined Growth Medium int | |
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