October 1, 2015 (Vol. 35, No. 17)

Kinase Selectivity Profiling Systems

Profiling kinase inhibitors is a critical step in the development of chemical probes or kinase drugs. The ability to generate an inhibitory profile of a compound across a broad collection of kinases is important for better understanding biological activity, avoiding any off-target effects, and in some cases, identifying new targets that could lead to novel therapeutic applications.

In the HTS phase, target-specific kinase assays are used to identify hit compounds. When profiling, robust universal assays are needed to assess the selectivity and potency of the inhibitor on multiple kinases from different classes. Most kinase profiling technologies suffer from drawbacks that limit their use in creating kinase inhibitor profiles. Limitations for in-house profiling include development and optimization of assays for each kinase in the panel, which can be costly and time consuming. Limitations associated with outsourcing compound profiling include confidentiality issues or result in the researcher not having immediate accessibility to the generated data.

Universal Assay Chemistry for Kinase Selectivity Profiling

The Kinase Selectivity Profiling Systems are easy-to-use systems that rely on the luminescent ADP-Glo™ Kinase Assay technology to characterize kinase selectivity. You get your results faster by profiling lead compounds in hours not days.

The ADP-Glo Kinase Assay is a universal platform for measuring kinase activity by quantifying the amount of ADP produced during a kinase reaction. We have optimized the ADP-Glo Assay technology with 174 Kinase Enzyme Systems spanning most of the human kinome. This optimization work allowed us to create the simple and convenient Kinase Selectivity Profiling Systems.

Each Kinase Selectivity Profiling System consists of a strip of eight enzymes with their corresponding substrates. Every kinase/substrate reaction is standardized for optimal kinase activity for inhibitor profiling. With these systems, you can perform selective inhibitor profiling with no assay development time and fewer resources. We provide sufficient material to profile up to 20 compounds at a single dose or to create a dose response for two compounds against eight kinases at once.

Simple Protocol for Flexible and Targeted Kinase Inhibitor Profiling

Reaction assembly is quick: Two simple dilutions provide working stocks of kinase and substrates, sufficient for 25 reactions. The one-time use design eliminates multiple freeze-thaw cycles and ensures optimal enzyme activity for each experiment. Formatted strips provide eight kinases per use, with the kinases organized either by kinase families to provide more relevant selectivity profiles or as a general panel containing 24 kinases spanning the human kinome to provide more comprehensive profiles.

Custom Kinase Selectivity Profiling Systems Meet Specific Research Needs

Use the web-based Custom Profiling Strips Selector Tool (www.promega.com/products/cell-signaling/kinase-assays/kinase-selectivity-profiling-in-next-
) to create a customized Kinase Selectivity Profiling System that matches your specific research needs.

The tool allows you to select the desired kinases in groups of eight to create a customized strip. Promega will automatically provide the optimized substrates for all kinases you select. Your custom Kinase Selectivity Profiling System uses all the same reagents as our standard Kinase Selectivity Profiling Kits:  the same ADP-Glo luminescent chemistry, the same optimized kinase reactions, and the same simple protocol. The custom systems specifically meet your research needs.


The Kinase Selectivity Profiling Systems combine the reliable ADP-Glo Assay chemistry and the expansive platform of complete Kinase Enzyme Systems to create easy-to-use, affordable systems for basic screening, kinase characterization, mode of action studies and selectivity profiling. With the Kinase Selectivity Profiling Systems, you rely less on third-party laboratories for profiling studies and get instant access to your profiling data, all without having to develop and optimize your own kinase assays. Furthermore, you have the option of creating your own customized profiling system to match your research needs.


Michael Curtin
Global Product Manager, 
Cellular Analysis and Proteomics
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