| Assay Method Information | |
| | Lipo-oxygenase Inhibitory Assay |
| Description: | Solution A was 50 mm DMAB in an 100 mm phosphate buffer (pH 7.0). Solution B was a mixture of 10 mm MBTH (3 mL) and hemoglobin (5 mg/mL, 3 mL) in 50 mm phosphatebuffer at pH 5.0 (25 mL). A linoleic acid solution was prepared by mixing 5 mg of linoleic acid with 50 mg Tween-20 in 3 mL water and then diluted with KOH 100 mmto a final volume of 5 mL. In the standard assay, the sample in ethanol (25 uL), SLO (EC 1,13,11,12; 4000 units/mL; 25 uL) and phosphate buffer, pH 7.0 (50 mm; 900 uL), were mixed in a test-tube and preincubation was carried out for 5 min at room temperature. A control test was done with the same volume of ethanol. Afterthe preincubation, linoleic acid solution (50 uL) was added to start the peroxidation reaction, and, 7 min later, solution A (270 uL) and then solution B (130 uL) were added to start the color formation. Further, 5 min later, 200 uL of a 2% SDS solution was added to terminate the reaction. |
| Affinity data for this assay | |
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