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| | Lipoxygenase Activity Assay |
| Description: | Lipoxygenase activity was measured in borate buffer solutions (0.1 M, pH 9) using the method described in the literature [Malterud et al., J. Agric. Food Chem., 48:5576-5580; Malterud et al., Pharmacology, 47:77-85], by measuring the absorbance at 234 nm for 60 s after addition of the enzyme (soybean 15-lipoxygenase) and linoleic acid (final concentration: 134 μM) as substrate at 20 ± 1°C. The final enzyme concentration was 167 U/mL. The synthesised substances were added as DMSO solutions (final inhibitor concentrations: 200, 100, 50, 25, 12.5, 6.25, 3.12 and 1.5 μM; final DMSO concentration 1%); whereas DMSO was added in the control experiments with no inhibitor. The mixture of each inhibitor and linoleic acid were set as the blank sample in the testing step. At least six control test tubes and three tubes for each inhibitor solution were measured. To ensure a constant enzyme activity throughout the experiment, the enzyme solution was kept on ice, and the controls were measured at regular intervals. |
| Affinity data for this assay | |
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