Solution Informationhelp
Enzyme: ATP-sensitive inward rectifier potassium channel 1
inhibitor: BDBM73214
substrate: n/a
 
Solution Type: Aqueous
pH at Preparation: n/a
Temp. Prep.: n/a
Comments: Experimental: Cells were plated in clear-bottomed, black-walled 384-well plates and cultured overnight in serum-free media containing Tet (for induction of ROMK expression) at 37 C in the presence of 5% CO2. The following day, the cells were combined with dye and incubated for 20min at RT, followed by automated washing using the ELx405 plate washer (BioTek, Winooski, VT). The plate was loaded into the Functional Drug Screening System 6000 (FDSS6000; Hamamatsu, Tokyo, Japan) to obtain an initial baseline image (F0) (10 images at 1 Hz; excitation, 470 +/- 20 nm; emission, 540 +/- 30 nm). DMSO vehicle control, TPNQ or test compound was added to each plate for final concentrations of 0.1 %, 2 uM and 10 uM, respectively. Four columns in each plate were filled with vehicle control or TPNQ-containing buffer for determination of Z', while the remaining 20 columns were used for compound screening at 10uM final concentration. The compound and cells were allowed to incubate for 20 min at RT,
 
 
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