| Assay Method Information | |
| | Enzyme Inhibition Assay |
| Description: | DPP4 activity was measured using a continuous fluorometric assay. The substrate, Gly-Pro-AMC, was cleaved by DPP4 to release the fluorescent AMC group. The reaction was monitored at the excitation wavelength of 360 nm and emission wavelength of 460 nm in a black 384-well plate using a PHERAstar Plus plate reader (BMG Labtech., Inc.) set at 37 C. The 30 ul assay solution contained 20 uM recombinant human DPP4 and 50 uM Gly-Pro-AMC substrate in assay buffer (100 mM HEPES, 100 mM NaCl, 0.1 mg/ml BSA, pH 7.5). The reaction was linear for at least 30 min (initial velocity) after addition of the substrate (no inhibitor control). All materials and reagents were pre-incubated at 37 C. prior to start of the experiment. Test compounds were serially diluted (3-fold) in 100% DMSO to avoid solubility issues; working solutions in 3% DMSO were prepared using assay buffer. 101 each of enzyme and compound solutions were mixed and pre-incubated at 37 C. for 30 minutes. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |