| Solution Information | help | |
| Enzyme: | Voltage-dependent T-type calcium channel subunit alpha-1H | |
| inhibitor: | BDBM76842 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | The purpose of this assay was to follow-up on the confirmatory and dose-response testing of the small molecule library provided by the Molecular Libraries Small Molecule Repository (MLSMR, distributed by DPI-Biofocus) for the ability to modulate calcium fluorescence in a Cav3.2 expressing cell line. This confirmatory assay tested 17 compounds chosen based on structure and favorable dose response results and reordered from multiple vendors in powder form for testing in 11-point 3-fold CRC experiments in triplicate with both the transfected (HH8) and untransfected cell lines (native HEK293). The native HEK cells contain a Ca2+ conductance that may be L-type and this was used as a first-tier selectivity screen. High-Throughput Screening Cells were plated in poly-D-lysine clear-bottomed, black-walled 384-well plates at 20,000/well and cultured overnight at 37 degrees C in the presence of 5% CO2. Cells were washed and loaded with Fluo-4 dye. The plates were loaded into the Hamamatsu FDSS | |
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