| Assay Method Information | |
| | 15-PGDH Enzyme Inhibition Test |
| Description: | A test compound and 4 nM recombinant human 15-PGDH (R&D Systems) were added, in 50 mM Tris-HCl (pH 8.0) containing 0.01% TWEEN 20 (Sigma) and 0.01% bovine gamma globulin (Sigma), to a 384-well Flat Bottom Black plate (Corning, 3820), and the resultant was allowed to stand still at room temperature for 12 minutes, and 30 μM PGE2 (Cayman Chemical) and 1 mM NAD+ (Sigma) were added thereto to start the reaction. 60 minutes after starting the reaction, measurement was performed using Synergy 2 (BioTeck) at an excitation wavelength of 340 nm and a fluorescence wavelength of 440 nm. Assuming that a fluorescence signal obtained with an assay buffer added instead of the test compound and NAD+ was 100% and that a signal obtained with NAD+ added was 0%, a concentration corresponding to 50% inhibition plotted on a concentration-reaction curve of the test compound was defined as an IC50 value |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |