| Solution Information | help | |
| Enzyme: | Voltage-dependent T-type calcium channel subunit alpha-1H | |
| inhibitor: | BDBM77087 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | The purpose of this assay was to test 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. All compounds were tested in single with a 10 point dose-response curve. 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 6000 and test compound added to give 10uM final concentration. After 20 minutes, the cells were stimulated with 5X Stimulation buffer (140 mM NaCl, 5 mM KCl, 1 mM MgCl2, 50 mM CaCl2, 5 mM glucose, 10 mM HEPES pH 7.3) and fluorescence measurements recorded. Data Analysis and Statistics The kinetic fluorescence values (F) from each well were divided by the initial frame of the read (F0) to give t | |
| If you find an error in this entry please send us an E-mail | ||