| Assay Method Information | |
| | In vitro Assay for Inhibition of DPP-4 |
| Description: | The DPP-4 Drug Discovery Kit (Enzo Life Sciences International, Inc.) was used for the assay of inhibition of DPP-4 activity. The assay is based on the cleavage of 7-amino-4-methylcoumarin (AMC) moiety from the C-terminus of the peptide substrate (H-Gly-Pro-AMC), which increases its fluorescence intensity at 460 nm. The DPP-4 inhibitor P32/98 was selected as a control. The substrate and DPP-4 enzyme were diluted 1/50 with assay buffer (50 mM Tris, pH = 7.5). 25 µL of assay buffer, 15 µL of enzyme solution and 10 µL of appropriately diluted solutions of the test compounds were added subsequently to 96-well microtitre plates. After incubation at 37 °C for 10 min, 50 µL of diluted substrate solution was added. Fluorescence was measured using an excitation wavelength of 380 nm and an emission wavelength of 460 nm by a Synergy H1 Multi-Mode Reader (BioTek, Beijing, China). |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |