| Assay Method Information | |
| | Fluorescence Resonance Energy Transfer (FRET) Assay |
| Description: | The activity of renin inhibitors in vitro was measured using the FRET assay, which was carried out in flat-bottom white opaque microtiter plates. The renin substrate in assay buffer was added to test compound in DMSO at various concentrations ranging from 10 uM to 1 nM. The trypsin-activated recombinant human renin was added to initiate the reaction. The increase in fuorescence at 495 nm (excitation at 340 nm) was measured for 60-360 min at room temperature using a Perkin-Elmer Fusion microplate reader. The slope of a linear portion of the plot of fuorescence increases as a function of time was then determined, and the rate was used to calculate % inhibition in relation to uninhibited control. The % inhibition values were plotted as a function of inhibitor concentration, and the IC50 was determined from a fit of this data to a four parameter equation. The IC50 is defined as the concentration of a particular inhibitor that reduces the formation of product by 50% relative to a control sample containing no inhibitor. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |