| Solution Information | help | |
| Enzyme: | Multidrug resistance-associated protein 1 | |
| inhibitor: | BDBM45156 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Cell Culture: H69AR cells (ATCC) were subcultured every 3-4 days in RPMI-1640 + 4.5 g/L glucose containing 10 mM HEPES pH 7.4 and 10% fetal bovine serum, and 2 mM L-glutamine (complete growth medium), incubated at 37 degrees C in 5% carbon dioxide, and maintained at sub-confluent density for no more than 20 passages. Compound Dosing/Plating: Dilution media, carrier and positive control drug were dispensed to dilution plate. Compounds were diluted in complete growth medium to prepare a 10X concentrated dosing solution (100 uM) with Biomek FX. 3.5 uL (1/10 final culture volume) of diluted compound was transferred to the cell plate (Corning part number:3712). Cell Plating: Twenty five uL of cells were dispensed (1250 cells per 25 uL complete growth medium) with 100 U/mL penicillin and 100 ug/mL streptomycin. Cells were maintained on ice during plating with stirring. Following plating, cell plates were briefly centrifuged to speed adhesion (< 1000 RPM for < 1 min), then incubated at r | |
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