Thrombin Human Chymotrypsin Serine Peptidase Enzymatic LeadHunter Assay - TW

Procedure Summary:

Human plasma thrombin is used. Test compound and/or vehicle is preincubated with 8.25 mU/ml enzyme‡ in modified HEPES pH 7.5 for 15 minutes at 25⁰C. The reaction is initiated by addition of 20 μM Z-Gly-Pro-Arg-AMC for another 60 minute incubation period. Determination of the amount of AMC formed is read spectrofluorimetrically at 360 nm/465 nm. Compounds are screened at 10 μM.

Substrate:

20 µM Z-Gly-Pro-Arg-AMC

Incubation:

60 min at 25°C

Control Inhibitor:

UNII-N62UL02WW4

Criteria For Significance:

≥ 50% of max stimulation or inhibition

Test Concentration / Dose:

Suggested concentration for initial testing is 10 uM

Test Sample Requirements:

Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).

Minimum Order Quantity:

2