| Assay Method Information | |
| | 15-PGDH Enzyme Inhibition Test |
| Description: | Test compounds and 4 nM recombinant human 15-PGDH (R&D systems) in 50 mM Tris-HCl (pH 8.0) containing 0.01% TWEEN 20 (Sigma) and 0.01% bovine gamma globulin (Sigma) were put into 384 Flat Bottom Black plates (Corning, 3820) and allowed to stand for 12 minutes at room temperature. Then, 30 μM PGE2 (Cayman chemical) and 1 mM NAD+ (Sigma) were added to start reaction. Sixty minutes after the start of reaction, signals were measured at an excitation wavelength of 340 nm and a fluorescence emission wavelength of 440 nm using Synergy 2 (BioTeck). The intensity of the fluorescent signal obtained when an assay buffer was added in place of the test compounds and NAD+ was defined as 100% and that obtained with the addition of NAD+ was defined as 0%. |
| Affinity data for this assay | |
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