Assay Method Information

Assay Name:  Enzyme Assay
Description:  Human DPP1 enzyme activity was determined by measuring release of the fluorophore aminomethyl coumarin (AMC) following enzymatic cleavage of the dipeptide substrate H-Gly-Arg-AMC. Assays were performed in black 384 well plates in 25 mM piperazine buffer, pH 5.0, containing 50 mM NaCl, 0.01% (v/v) Triton X100, 5 mM DTT with 0.35 nM human DPP1 enzyme, 300 μM H-Gly-Arg-AMC substrate (˜Km concentration) and test compounds at 0.51-10000 nM concentration range. Human DPP1 was pre-incubated with test compounds for 30 minutes at 25° C. prior to substrate incubation for a further 40 minutes at 25° C. Enzyme activity was determined by measuring fluorescence of the AMC product at excitation and emission wavelengths of 380 nm and 460 nm using a BMG LABTECH PHERA star plate reader.
Affinity data for this assay
 

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