| Solution Information | help | |
| Enzyme: | Mitochondrial import inner membrane translocase subunit TIM23 | |
| inhibitor: | BDBM78194 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Materials: Yeast Strain: ySHDSTim23ts (tim23-1 strain) Maintenance agar: YPD agar (Teknova, # Y1001) Growth Medium: YPD broth (Teknova # Y5000) SD Assay Medium: 1.2 g/L Yeast Nitrogen Base w/o amino acids, 5 g/L Ammonium Sulfate, 20 g/L Dextrose, 13.8 g/L Succinate (Sunrise Science Products, # 1998-046-1L), supplemented with 2 g/L SC Amino Acid Mix (MP Biomedicals, # 4400-022)# SD Minimal Medium: 1.2 g/L Yeast Nitrogen Base w/o amino acids, 5 g/L Ammonium Sulfate, 13.8 g/L Succinate, (Sunrise Science Products, # 1998-047-250ml) Assay Plate: Corning 1536 Well White Plate (# 3725) Detection Reagent: BacTiter-Glo, (Promega, # G8233) I. Compound Addition: 1. Using LabCyte Echo 555, transfer 40 nL of DMSO to positive and negative control wells in columns 1 - 4. Using a dose response protocol, transfer compounds from 10 mM and 0.312 mM Echo qualified plates into assay plate columns 5 - 46. (Final concentrations range 100 uM to 0.2 uM, 10 doses, in triplicate with 1% DMSO.) 2. Centri | |
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