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GYRASOL TECHNOLOGY PLATFORM Electron Transfer Quenching No requirement for spectral overlap between donor and acceptor molecules Advantages • Quench of all commercial fluors • Simultaneous monitoring of several fluor-labeled substrates Performance Summary •Uses standard fluorescent instrumentation •Direct detection of substrate conversion •Homogeneous one-step "mix and measure" •No radioactivity or antibodies •Linear dose response •Easily scaled and automated for HTS •High Substrate Tolerance (1 μM -100 μM) •High ATP tolerance (up to 1 mM) •Adaptable to a variety of substrates: Lipids, peptides, oligonucleotides and cyclic nucleotide •Multiplexable •Precise quantification of product conversion using calibrators •Stable detection product (~24h) •Cost effective TECHNOLOGY DESCRIPTION The Gyrasol Technology platform directly measures the activity of kinases, phosphatases, phosphodiesterases and proteases. The platform is robust, homogeneous and adaptable to standard instrumentation. The Gyrasol Sensor is a proprietary small molecule that contains a trivalent metal ion, which binds to phosphoryl groups on biological substrates. Suitable substrates include peptides, lipids, oligonucleotides and cyclic nucleotides that can be labeled with any fluor. The presence or absence of substrate phosphoryl groups is measured by the change of fluorescence of the fluor-labeled substrate when bound by the Sensor. The change in fluorescence directly correlates to the level of substrate conversion (Figure 1).  Figure 1: Schematic depicting a Gyrasol assay A substrate labeled with a fluorescent dye is reacted with target enzyme resulting in substrate with or without phosphoryl groups. The fluorescence of the substrate is quenched when the Gyrasol Sensor (yellow star) associates to phosphoryl moieties on the substrate. |
