| Solution Information | help | |
| Enzyme: | 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase beta-3 | |
| inhibitor: | BDBM114297 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this biochemical assay is to determine PLC-B3 inhibitory dose response curves of compounds that confirm activity in a set of previous experiments entitled: "Fluorescence-based biochemical high throughput confirmation assay to identify inhibitors of phospholipase C isozymes (PLC-B3)." (AID 743261). In this assay, PLC-B3 isozyme is incubated with test compounds and fluorogenic reporter WH-15. As designed, test compounds that act as PLC-B3 inhibitors will prevent the hydrolysis of WH-15 fluorogenic reporter, thus preventing the release of IP3, a quinomethide derivative, and 6-aminoquinoline, which is highly fluorescent, leading to decreasing well fluorescence. Compounds are tested in triplicate using a 10-point 1:3 dilution series starting at a maximum nomimal test concentration of 122.5 uM. (SIDs 14730721, 14742430, 24788424, 24809205, 24841228, 26663487, 49665443, 50085435, 92764458, 93576637, and 93577326 were tested as sextuplicates.) Protocol Summary | |
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