IMAP Enzyme Assays

Enzymes involved in cellular signaling include kinases, phosphatases, and phosphodiesterases (PDEs). Protein kinases, which modify other proteins by chemically adding phosphate groups to them, comprise one of the largest classes of molecules targeted by drug discovery programs. Deregulation of kinase expression and/or function can result in cancers and other diseases. Protein phosphatases and PDEs are key regulators of signal transduction pathways that are disrupted in cancers, neurodegeneration, diabetes, and others disorders.

Until recently, researchers performed assays for kinase, phosphatase, and PDE activity using radioactive isotopes, which pose safety and disposal challenges, or highly specific antibodies, whose development is costly and time-consuming. To address these issues, Molecular Devices[MS1]  introduced its proprietary IMAP® technology, a non-radioactive, homogeneous assay that is based on detection of phosphogroups and is thus a generic platform for accurate determination of kinase, phosphatase, and PDE activity. Enzyme activity can be measured using either fluorescence polarization or time-resolved FRET detection. To assist researchers in choosing the appropriate substrate for their kinase of interest, Molecular Devices also offers IMAP Substrate Finder Kits, allowing researchers to quickly test one or more kinases against dozens of peptides included in a 384-well microtiter plate.

To simplify assay development and deliver maximum performance, the IMAP Assay Kits have been optimized for use with Molecular Devices' FlexStation® 3 and SpectraMax® M4 and M5/M5e Systems. The kits are conveniently packaged in bulk kits for high throughput screening applications and Explorer kits for lower throughput applications.

Protein kinases are enzymes that modify other proteins by chemically adding phosphate groups to them. Cellular signaling through kinases controls cellular growth, movement, and death; therefore, kinases are highly regulated in cells. Deregulation of kinase activity can result in cancer and other diseases. Kinases are one of the largest drug targets, along with GPCRs, and kinase inhibitors are currently being developed as drugs such as Gleevec®, a kinase inhibitor currently in clinical use.

Phosphatases are enzymes that remove phosphate groups from their substrates. Enzymes can be activated or deactivated by the addition of phosphate groups, so phosphatases are important to many cellular signaling pathways. Deregulation of phosphatase activity is involved in a variety of disorders including neurodegeneration, cancer, diabetes, and obesity.

IMAP Substrate Finder Kit assists researchers in choosing the appropriate substrate for the kinase of interest. The IMAP FP Substrate Finder Kit enables the researcher to quickly test one or more kinases against dozens of peptides included in a 384-well microtiter plate to identify potential substrates for subsequent IMAP screens.

Cyclic nucleotide phosphodiesterases (PDEs) are enzymes that degrade the phosphodiester bond of the second messenger molecules cAMP and cGMP, and are therefore key regulators of cellular signal transduction mediated by these molecules. PDE inhibitors are being investigated as potential therapeutics for pulmonary disease, coronary heart disease, depression, dementia, and schizophrenia. The currently available drugs Viagra® and Pletal are inhibitors of PDE activity.

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