| Assay Method Information | |
| | PREP and DPP4 Protease Activity Assay |
| Description: | PREP activity was measured with recombinant human PREP (R&D systems, #4308-SE). The PREP activity assay was performed using the FACS buffer (PBS including 1% FBS, Sigma-Aldrich, Cat #D8537). As substrate 50 μM Z-GP-AMC (Bachem, #4002518) was used. The DPP4 activity assay was performed in DPP assay buffer (25 mM Tris, pH 8.0). Recombinant human DPP4 was purchased from R&D systems (#9168-SE). 20 μM of GP-AMC (Santa Cruz Biotechnology, #115035-46-6) was used as substrate.Fluorescence of AMC (excitation at 380 nm and emission at 460 nm) after cleavage was measured in a kinetic mode for 5 minutes at 37° C. in a SPECTRAmax M5 plate reader. RFU/sec was calculated by SoftMax Pro software and plotted against test compound concentration. Four parameter logistic (4PL) curve fitting and pIC50 calculations were performed using ActivityBase software. |
| Affinity data for this assay | |
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