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| Solution Information | help | |
| Enzyme: | ATP-dependent molecular chaperone HSP82 | |
| inhibitor: | BDBM54164 | |
| 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 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) Pen/Strep Fungal Inoculum Test Strain: C. albicans CaCi-2 Inoculate 500 ul of strain from cryopreserved stock into 250 ml shaker flask containing 30 ml YPD medium. Shake at 30C O/N. Spin down culture, pour off broth, wash with RPMI medium. Spin down, pour off broth, resuspend in RPMI medium. Read OD 600 of fungal culture using standard plate reader. Dilute to starting OD of the fungal inoculum of 0.00015 A600. Prepare Geldanamycin positive control solution: 1 ml 15mM stock solution Geldanamycin 4 ml RPMI Growth Medium 50 ul Pen/Strep Dispense 400 nL into positive control wells using Combi NL (Thermo.) Dispense 20 uL/well of RPMI synthetic defined medium into 384-well black plates. Pin 100 nL from compound | |
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