| Solution Information | help | |
| Enzyme: | Tropomyosin alpha-1 chain | |
| inhibitor: | BDBM52817 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this assay is to determine dose response curves for compounds that confirmed activity in a set of previous experiments entitled, "Fluorescence-based biochemical high throughput confirmation assay for inhibitors of the calcium sensitivity of cardiac Regulated Thin Filaments (RTF)" (AID 504635). This biochemical assay employs the visible fluorophore, IANBD (Ex ~ 485 nm; Em ~ 535 nm) attached to troponin C (TnC, the Ca2+ binding subunit of the RTF) to monitor the Ca2+ dependent changes in fluorescence arising from IANBD-labeled RTF. As designed, a decrease in the RTF fluorescence intensity at a fixed calcium concentration and wavelength in response to a test compound will indicate that a change in the RTF calcium sensitivity has occurred. This assay employs an EC70 calcium concentration (1.6 mM). Compounds are tested in triplicate using a 10-point, 1:3 dilution series starting at a nominal concentration of 71.5 uM. Protocol Summary: Prior to the start of | |
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