Abstract
Tracy Worzella1, Brad Larson1, Kevin Hsiao1 and Aoife Gallagher2
1Promega Corporation, 2800 Woods Hollow Road, Madison WI 53711, USA
2Deerac Fluidics, Unit 8 Enterprise Centre, Pearse Street, Dublin 2, Ireland
We demonstrate the use of Promega Kinase-Glo® Plus Luminescent Kinase Assay for miniaturized kinase assays using Deerac Fluidics Equator™ HTS liquid handling system. The Kinase-Glo® Plus Assay is a homogeneous method of measuring kinase activity by quantifying ATP in solution following a kinase reaction. The Equator™ HTS is a noncontact liquid dispenser capable of delivering volumes from 20μl down to 50nl.
Because assay development and screening are key uses for miniaturized assays, we show the Equator™ HTS can be used to optimize Kinase-Glo® reactions and screen for kinase activity. We use the Equator™ HTS to titrate kinases and substrates to determine optimal concentrations for subsequent screening activities in 1536-well format. A panel of kinases and compounds are counterscreened for kinase selectivity profiling. 24-point serial titration is performed to determine potency. Data show that miniaturization of the Kinase-Glo® Plus Assay does not compromise assay quality. The combination of Kinase-Glo® Plus and the Equator™ HTS provides an integrated solution for kinase drug discovery.