Notes: The authors compared different in vitro assay formats, including viable cells, cell lysates and recombinant enzymes for their ability to identify MAO inhibitors. The assays used in this paper were the fluorescent kynuramine assay (recombinant enzyme and live cell), a fluorescent Ampliflu Red assay (cell lysate), and the luminescent MAO-Glo™ Assay (recombinant enzyme). The fluorescent and luminescent assays were found to be in good agreement regarding human recombinant MAO A and B. (4521)

Notes: The authors describe the modification of the Promega MAO-Glo™ Assay to create a robust high-throughput screen for semicarbazide-sensitive amine oxidases (SSAOs). These proteins are implicated in inflammatory diseases and are believed to play a role in immune cell trafficking and cell activation. Their modified MAO-Glo™ Assay was tested with enzyme from recombinant and cell sources, and was able to identify specific inhibitors. (4155)

Notes: The authors of this review article highlight the use of bioluminescence as a readout for high-throughput ADME/Tox assays. They discuss three strategies for designing bioluminescent assays, using either luciferase, ATP or luciferin substrates as the limiting reagents for a luciferase-catalyzed reaction. Reporter gene assays limit the production of luciferase by tying it to a promoter or DNA regulatory region of interest. Such assays can be used to study genes that are regulated by drugs and other xenobiotics. Bioluminescent assays in which ATP is the limiting reagent of the luciferase reaction can be designed to monitor cell viability or the activity kinases. Bioluminescent assays in which the substrate is limiting can be designed so that the activity of a particular enzyme results in the production of a luciferin substrate that can, in turn, be acted upon by luciferase. (3926)

Notes: The authors of this paper review bioluminescent assay technologies, discussing HTS reporter, cell-based and luciferase biosensor assays. They divide luminescent assays into three basic categories: assays that measure ATP concentration (cell viability and kinase assays), assays that measure changes in luciferase levels (reporter assays, GPCR assays), and assays that measure changes in luciferin levels (protease [including caspase], P450 and MAO assays). (3737)