| Assay Method Information | |
| | Human Neutrophil Elastase Inhibition Assay |
| Description: | HTS was performed in black flat-bottom 96-well microtiter plates. The reaction was initiated by addition of elastase substrate to the reaction buffer containing enzyme and test compounds. Kinetic measurements were obtained using a Fluoroskan Ascent FL fluorescence microplate reader (Thermo Electron, MA) with excitation and emission wavelengths at 355 and 460 nm, respectively. For selected lead compounds, the inhibition constant (Ki) values were determined using Dixon plots of three to four different concentrations of the substrate. At each substrate concentration, rates were determined with four to five different inhibitor concentrations, and the inverse of the velocities was plotted against the final inhibitor concentration. Ki was determined from the intersection of the plotted lines. |
| Affinity data for this assay | |
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