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A Simple and Robust Automated Kinase Profiling Platform using Luminescent ADP Accumulation Technology下載
August 13, 2009
Authors: Brad Larson and Peter Banks, BioTek Instruments, Winooski, VT, Hicham Zegzouti and Said Goueli, Promega Corporation, Madison, WI
Kinases continue to be an important target class in today’s drug discovery efforts. Following the identifi cation of lead compounds through screening efforts, it is important to profi le these leads against other kinases to ascertain potential off-target effects. Because different kinases included in a profi le require the use of different buffers, substrates, ATP, and incubation times, the proper choice of assay and instrumentation is essential to minimize optimization time and cost. Here we demonstrate the versatility of a luminescent ADP accumulation assay, where one set of reagents is used for kinases with differing ATPKm app. We have used a simple, inexpensive automated pipetting system to automate the entire process in 384-well format, from assay optimization through generation of compound IC50 values. Agreement with literature values proves this combination of chemistry and instrumentation provides a simple, yet robust solution for automated kinase profi ling. Kinases are one of the most diverse enzyme families being studied today.