| Assay Method Information | |
| | Inhibition of DPP1 Enzyme Activity Assay |
| Description: | Experimental Methods1) Preparation and handling of compounds: Accurately weigh the compound and dissolve it in 10 mL of DMSO to prepare a 10-mL of the bulk, which is further diluted as required.2) Screening test: Dilute recombinant human cathepsin rhCathepsin C/DPP1 and recombinant human cathepsin rhCathepsin L with an activating buffer solution, and incubate at 37° C. for 60 min. Add 4 μL of test compound to the white well of the 384-well plate. Add 4 μL of the enzyme mixture after incubation to the white wells. Block the 384-well plate and allow to stand at room temperature for 30 min, and add 8 μL of 2-fold diluted Gly-Arg-AMC (hydrochloride). Block the 384-well plate and allow to stand at room temperature for 2 h. Prepare 4× stop solution and add 4 μL stop solution to white wells. Read fluorescence data on the fluorescence meter Victor Nivo35.3) Data analysis:All IC50 values are converted to percent inhibition using Prism Graphpad 8.0. |
| Affinity data for this assay | |
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