Jewess P J, Devonshire A L
Department of Biological and Ecological Chemistry, IACR-Rothamsted, Harpenden, Hertfordshire, AL5 2JQ, United Kingdom.
Anal Biochem. 1999 Jul 15;272(1):56-63. doi: 10.1006/abio.1999.4168.
Kinetic microplate-based assays for both mitochondrial NADH:ubiquinone oxidoreductase (complex I) and succinate:cytochrome c oxidoreductase using insect submitochondrial particles as the source of the enzyme activities have been developed. These assays have been used to design high-throughput screens for inhibitors of these mitochondrial electron transfer activities to assess their intrinsic in vitro efficacies as potential pesticides. These methods can be used to test up to 60 compounds per day without the use of automated sample handling and diluting technology. The accuracy, specificity, and reproducibility of the microplate methods compared well with conventional spectrophotometer-based assays.
已开发出基于微孔板的动力学测定方法，用于线粒体NADH：泛醌氧化还原酶（复合体I）和琥珀酸：细胞色素c氧化还原酶，使用昆虫亚线粒体颗粒作为酶活性来源。这些测定方法已用于设计高通量筛选，以筛选这些线粒体电子传递活性的抑制剂，以评估它们作为潜在农药的内在体外效力。这些方法可用于每天测试多达60种化合物，无需使用自动样品处理和稀释技术。与传统的基于分光光度计的测定方法相比，微孔板方法的准确性、特异性和可重复性良好。
