| Assay Method Information | |
| | Inhibition Assay |
| Description: | To activate the proenzyme, 5 ul procathepsin K were mixed with 1 ul activation buffer, and incubated at room temperature for 30 min.5 uL test compound (final concentration 0.1 nM to 100 uM) in aqua bidest (containing 4% DMSO, final DMSO concentration 1%) were mixed with 10 uL of Cathepsin K in assay buffer (final concentration 2 ng/uL) and incubated for 10 min. Then 5 uL of substrate in assay buffer (final concentration 12.5 uM) were added. The plates were then incubated at room temperature for 60 min. Then, the reaction was stopped by adding 10 uL of E64 in assay buffer (final concentration 1 uM). The fluorescence in the wells was determined using a Molecular Devices SpectraMax M5 Fluorescence Reader (Ex 360 nm, Em 460 nm).Each assay microtiter plate contains wells with vehicle controls (1% DMSO in bidest) as reference for non-inhibited enzyme activity (100% Ctl; high values) and wells with inhibitor (E64 in bidest+1% DMSO. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |