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Assay Method Information |
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| | Inhibition Assay |
| Description: | Each of Compounds (1) to (7) was added into 50 μL buffer A (containing 20 mM Tris-HCl (pH 7.4), 0.1% NaN3 and 5 mM CaCl2) to obtain compound solutions with concentrations of 0.02, 0.2, 2, or 20 μM for each of Compounds (1) to (7), followed by m adding 50 μL of the compound solutions into wells of a 96-well plate. The buffer A and Sivelestat (under the same solvent condition and concentrations as those of Compounds (1) to (7)) with the same volume were respectively used as a control group and a positive control group. Each well of the 96-well plate was further added with 25 μL of human neutrophil elastase (Enzo, 200 nM, in buffer B containing 20 mM Tris-HCl (pH 7.4) and 0.1% NaN3) and 25 μL MeO-Suc-Ala-Ala-Pro-Val-p-nitroanilide (500 μM, prepared in the buffer B), followed by reacting in an incubator (30° C.) for 30 minutes. |
| Affinity data for this assay | |
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