| Assay Method Information | |
| | In vitro DPP IV enzyme inhibition assay |
| Description: | The enzyme assay was performed using DPP IV drug discovery kit (BML-AK 499; Enzo Life Sciences, Plymouth Meeting, PA, USA). The activity of test compounds was assayed using human recombinant DPP IV enzyme, chromogenic substrate (H-Gly-Pro-AMC, Km 114 µM), DPP IV inhibitor (P32/98), assay buffer and calibration standard as provided in the kit. Following the manufacturer's guidelines, the assay was performed using 96-well flat-bottomed microtitre plate followed by addition of assay buffer, DPP IV enzyme and chromogenic substrate (HGly-Pro-pNA). Test compounds were dissolved in DMSO at different concentrations of 25, 50, 100, 200 µg/mL, and 20 µL were added to each well after further dilutions. The plate was incubated at 37°C for 10 min and read continuously at A-405 nm. |
| Affinity data for this assay | |
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