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Assay Method Information

Assay Name:  Protease Inhibition Assay
Description:  The inhibition of plasma kallikrein and coagulation factor FXII was assessed by incubating the proteases with bicyclic peptide (twofold dilutions) and measuring the residual protease activity with fluorogenic substrates. The inhibition assays were performed in buffer (150 mL) containing Tris·HCl (pH 7.4, 10 mm), NaCl (150 mm), MgCl2 (10 mm), CaCl2 (1 mm), bovine serum albumin (BSA, 0.1 %, w/v), Triton-X100 (0.01%, v/v), and DMSO (5 %, v/v). Final concentrations of human plasma kallikrein (Innovative Research) and human FXII (Innovative Research)were 0.25 nm and 10 nm, respectively. The fluorogenic substrates Z-Phe-Arg-AMC (plasma kallikrein) and Z-Gly-Gly-Arg-AMC (FXII) were used at a final concentration of 50 mm. Fluorescence intensity was measured with a Tecan Infinite M 200 Pro plate reader (λex= 368 nm, λem=468 nm).
Affinity data for this assay
 

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Last update November 1, 2007
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