| Solution Information | help | |
| Enzyme: | Tumor necrosis factor ligand superfamily member 10 | |
| inhibitor: | BDBM44373 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | 1) MDA-MB-435 cells and TRAIL were provided by the assay provider 2) ATPLite (Perkin Elmer) Cytotoxicity assay protocol: Day 1 1) MDA-MB-435 cells are grown in DMEM, 10% FBS, 4.5 g/l glucose, 2mM L-glutamine, 1mM Na-pyruvate, 15ug/ml gentamycin and are harvested at 80-90% confluency. 2) Cells are then seeded into a 384 well plate (Greiner # 781098 white, clear-bottom, 384, TC-treated plate) at a concentration of 500 cells/well in 45ul and incubated for 24 hours at 37 oC in a CO2 incubator to obtain 10-20% of confluence. Day 2 1) Add 5 ul of 20X compound in 10% DMSO to wells to obtain 10 uM (final assay concentration of DMSO is 0.5% in 100ul final assay volume per well). *Drugs are tested in 10-point dilution series with dilution factor 2 starting from 10uM final assay concentration. 2) Add 45uL of Assay Media to each well 3) Incubate cells 4 hours at 37 oC in a CO2 incubator. 4) Add 5 ul of 20X TRAIL in PBS without Mg2+ and Ca2+ (Final assay concentration of TRAIL is 100ng/mL in | |
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