| Assay Method Information | |
| | Inhibition Assay |
| Description: | The test was carried out according to the method of Urade, Y. et al. (J. Biol. Chem., 262, 3820-3825, (1987)). More specifically, the reaction mixture (49 uL) containing 100 mM Tris-HCl (pH 8.0), 1 mM reduced glutathione, 0.1 mg/mL gamma-globulin, and human H-PGDS (q.s.), and a compound (final concentration: 0.01-100 uM) was preincubated at 25 C. for 5 minutes. Note that a DMSO solution (final concentration: 1%) was added to the solvent control group. Subsequently, 1 uL of [14C] prostaglandin H2 (final concentration: 10 uM) was added to start the reaction. One minute after the start of the reaction, 250 uL of a reaction stopper solution (diethylether/methanol/1 M citric acid (30/4/1) at a temperature of -20 C. was added to stop the reaction. After the reaction was stopped, 50 uL of the upper layer portion (organic solvent phase) was applied to a TLC plate and developed at -20 C. for 45 minutes. |
| Affinity data for this assay | |
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