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Promega Corporation

Profiling Kinase Activities Using a Universal Luminescent ADP Detection Platform...

Profiling Kinase Activities Using a Universal Luminescent ADP Detection Platform Scientific Poster

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Abstract

Hicham Zegzouti, Jolanta Vidugirience, Kevin Hsiao, Mary Sobol,  Dongping Ma and Said Goueli
Promega Corporation, Madison, WI

Because the selectivity of a kinase inhibitor towards the target of interest is critical for its advancement through the various phases of drug development, profiling of the inhibitor against other kinases is a common practice in drug discovery. Thus, a robust  and universal kinase assay is desired in order to assess the selectivity and potency of the inhibitor on multiple kinases from different classes that often use substrates with different chemical structures. Radiometric assays have been widely accepted as the most reliable platform for profiling kinases. Although several other technologies were developed in the last few years, most suffer from a variety of limitations that make it difficult to address al the needs of kinase screening, mode of action (MOA) studies and profiling with a single platform during the  drug discovery process. ADP-Glo™ technology measures kinase activity by quantifying the amount of ADP produced during the reaction. This assay is universal, applicable to all kinds of kinase substrates regardless of their nature with no prior modification (peptides, proteins, alcohols, lipids and sugars) making it easier to create inhibitor profiles of different kinase families using one platform. The assay is as sensitive as a radiometric method but without the use of radioactivity and disposal of radioactive waste. Because of the high signal to background (S/B) values even at low % ATP to ADP conversion the assay allows significant savings of enzyme usage in kinase assays. The fact that ADP-Glo™ assay offers so many positive attributes makes it an ideal assay not only for primary and secondary screening but also for profiling of lead compounds.

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  • Part# PS104
  • Printed in USA.

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