Kinase Profiling — Simplified

Kinase screening is a crucial, but costly, necessity for developing targeted kinase inhibitors. To simplify kinase profiling, Promega offers:

  • The Kinase Selectivity Profiling Systems, which include kinase and substrate pairs grouped in single kinase families or as a general panel of kinases representative of the human kinome for a broad kinase profile.
  • The Kinase Enzyme Systems, which provide the enzyme, substrate, buffer and ADP-Glo™ reagents in an integrated, easy-to-use protocol for quantifying the activity of over 170 kinases, including lipid kinases.

ADP-Glo™ Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is converted into light by Ultra-Glo™ Luciferase. The luminescent signal positively correlates with kinase activity. The assay is well suited for measuring the effects of chemical compounds on the activity of a broad range of purified kinases, making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo™ Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP.

The Kinase Selectivity Profiling Systems are optimized for fast and simple kinase profiling reactions and include kinase and substrate pairs grouped in single kinase families or as a general panel of kinases representative of the human kinome for a broad kinase profile. These systems rely on the proven ADP-Glo™ Kinase Assay technology to make kinase profiling fast and simple.

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An overview of the Kinase Selectivity Profiling System protocol. Concentrated kinases and substrates/cofactor stocks are first diluted in the 8-tube strip, then 2µl of each is combined with 1µl of test compound in a 384-well plate. After a 1-hour incubation, the ADP-Glo™ Assay is performed. The resulting luminescent signal is proportional to ADP concentration and correlates with kinase activity.

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An overview of the Kinase Selectivity Profiling System protocol. Concentrated kinases and substrates/cofactor stocks are first diluted in the 8-tube strip, then 2µl of each is combined with 1µl of test compound in a 384-well plate. After a 1-hour incubation, the ADP-Glo™ Assay is performed. The resulting luminescent signal is proportional to ADP concentration and correlates with kinase activity.

Each Kinase Enzyme System is comprised of a recombinant human kinase, substrate, buffer and supplemental reagents optimized for use with the ADP-Glo™ Kinase Assay. An Application Note for each KES is available to provide specific information on how to use the reagents with the ADP-Glo™ Kinase Assay and minimize the optimization required.

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Promega lipid kinase enzymes, substrates and detection systems provide a complete set of reagents for performing phosphoinositide lipid kinase (PIK) reactions using a luminescent ADP-detection platform, the ADP-Glo™ Kinase Assay. The reagents include purified human recombinant proteins of Class I PI3Ks, optimized reaction buffer and ready-to-use lipid kinase substrates.

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