| Assay Method Information | |
| | 15LO Inhibition Assay |
| Description: | Linoleic acid and two assay solutions (A and B) were prepared in advance. Solution A was 50 mM 3-(dimethylamino)benzoic acid (DMAB) in a 100 mM phosphate buffer (pH 7.0). Solution B was a mixture of 10 mM 3-methyl-2-benzothiazolinone (MBTH) 3 mL, hemoglobin (5 mg/mL, 3 mL) in 50 mM phosphate buffer 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 diluting with KOH 100 mM to a final volume of 5 mL. In the standard assay, the sample in ethanol (25 mL), soybean 15-lipo-oxygenase (SLO) (4000 units/mL in 50 mM phosphate buffer pH 7.0; 25 mL) and phosphate buffer pH 7.0 (50 mM; 900 mL) 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. After the preincubation, linoleic acid solution (50 mL) was added to start the peroxidation reaction, and 7 min later,solution A (270 mL) and then solution B (130 mL) was added to |
| Affinity data for this assay | |
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