| Solution Information | help | |
| Enzyme: | G protein-activated inward rectifier potassium channel 1 | |
| inhibitor: | BDBM50004233 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Protocol: HEK293 cells expressing alpha2C/GIRK were plated at 15,000 cells per well in Dulbecco's modified medium (DMEM) containing 10% dialyzed fetal bovine serum (FBS), 100units/mL penicillin/streptomycin, and 20mM HEPES. 384-well, black walled, clear bottom, poly-D-lysine coated plates were used. After overnight incubation at 37degreesC in the presence of 5% CO2, the medium was removed and BTC-AM (Molecular Probes) dye in assay buffer (Hanks Balanced Salt Solution (HBSS)) was added. After 1hour, the dye was replaced with HBSS. Test compounds were prepared in thallium assay buffer (125mM sodium gluconate, 1mM magnesium sulfate, 1.8mM calcium gluconate, 5mM glucose, 12mM thallium sulfate, 10mM HEPES, pH 7.3). The compounds were added to the cell plate by the Hamamatsu FDSS. Each cell plate was imaged at 1Hz for 10seconds, 10uL of test compound was added, and the cell plate was imaged at 1Hz for 2minutes total sampling time. Data Processing: Each kinetic trace was normalize | |
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