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Urokinase Inhibitor Screening Service
Summary
Urokinase (uPA, Urokinase plasminogen activator) (3.4.21.31) is a key serine protease involved in the degradation of the extracellular matrix that catalyzes the conversion of plasminogen to active plasmin. It acts as a thrombolytic agent to break up blood clots and when over-expressed, has been reported to influence the growth of certain malignant tumors (breast, prostate, etc).
Amiloride Hydrochloride IC50 Determination
To test the suitability of BioAssay System’s Urokinase Assay for screening for inhibitors, we performed an initial experiment with a known Urokinase inhibitor, amiloride hydrochloride, at 0.75 U/µL human Urokinase. Using 0.5 mM Z-GGR-AMC substrate, we observed an IC50 of 16.6 µM for amiloride hydrochloride. The percent enzyme activity was plotted versus the concentration of amiloride hydrochloride. This value is close to the reported 7 to 12 µM IC50 for this inhibitor (https://pubchem.ncbi.nlm.nih.gov/bioassay/1265722).
Key Features
To test the suitability of BioAssay System’s Urokinase Assay for screening for inhibitors, we performed an initial experiment with a known Urokinase inhibitor, amiloride hydrochloride, at 0.75 U/µL human Urokinase. Using 0.5 mM Z-GGR-AMC substrate, we observed an IC50 of 16.6 µM for amiloride hydrochloride. The percent enzyme activity was plotted versus the concentration of amiloride hydrochloride. This value is close to the reported 7 to 12 µM IC50 for this inhibitor (https://pubchem.ncbi.nlm.nih.gov/bioassay/1265722).
Key Features
- High-throughtput. Assays are carried out in 96/384-well plate.
- Robust. Has a Z’ factor of >0.56.
- The assay tolerates up to 0.3 v% DMSO.
- Urokinase inhibitor hit profiling.
- Urokinase inhibitor lead generation.
- Structure-activity relationship studies.
