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Assay Method Information |
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| | Monoamine Oxidase Assay |
| Description: | A fluorimetric method reported by Matsumoto et al.[T. Matsumoto, O. Suzuki, T. Furuta, M. Asai, Y. Kurokawa, Y. Nimura, Y. Katsumata, I. Takahashi. Clin. Biochem. 1985, 18, 126.] was used to determine MAO-A and MAO-B activities. The substrate and mitochondrial fractions were incubated for 30 min at 38 °C followed by the inhibition of one of the MAO isoforms with the particular inhibitor (L-deprenyl (10.5 mm) to determineMAO-A activity and clorgyline (10.5 mm) to determine MAO-B activity). Propylene glycol was used to dissolve the incubation mixture containing compounds at five different concentrations in the range of 0.5 nm to 0.1 m, specific substrate for MAO-A or MAO-B (0.1 mm), mitochondrial suspension (6 mg/mL), and 0.1 mL phosphate buffer (0.25 m, pH 7.4). In a shaking water bath, the mixture was incubated for 60 min at 37 °C. Perchloric acid was added to quench the reaction. This was followed by the centrifugation of the samples at 10 000 g for 5 min followed by the collection of 2.7 mL supernatant using 1 N NaOH. Shimadzu RF-6000 Spectrofluorimeter was used to record the measurements. |
| Affinity data for this assay | |
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