| Assay Method Information | |
| | Elastase inhibition assay |
| Description: | Serially diluted compounds in DMSO were dispensed into a 384-well black opaque plate by Echo dispenser. 0.1 μg/mL human neutrophil elastase (EPC, Catalog# SE563, Owensville, Ms.) or human sputum diluted with assay buffer (100 mM HEPES, 500 mM NaCl, 0.02% Tween 20) was added into the 384-well plate, and was incubated with different compounds at different concentrations for 30 minutes at room temperature. The final concentration of DMSO in the reaction was 0.1%. Elastase substrate MeOSuc-AAPV-AMC (Bachem, Catalog #I-1270, Torrance, CA) of 100 μM final concentration was then added into the reaction system just before enzyme kinetics were read on PheraSTAR plate reader at excitation of 380 nm and emission of 460 nm with a 3-minutes interval for 30 minutes in total. |
| Affinity data for this assay | |
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