The generation of H 2O 2 was dependent on the concentration of both the compound and DTT and was abolished by catalase. The phenol red-horseradish peroxidase (HRP) assay readily detected H 2O 2 either added exogenously or generated by the redox cycling of compounds in dithiothreitol (DTT). We report here the development and optimization of a simple 384-well colorimetric assay to measure H 2O 2 generated by the redox cycling of compounds incubated with reducing agents in high-throughput screening (HTS) assay buffers.
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