A valuable tool for kinase drug discovery is an enzyme assay that measures catalytic function. The functional assay can be used to identify inhibitors, estimate affinity, characterize molecular mechanisms of action (MMOAs) and evaluate selectivity.

2012年5月1日 · The keys to protein kinase assay development lie in the ability to 1) choose an appropriate “readout” technology, 2) have ample quantities of enzymes, cell lines, antibodies and reference compounds, and 3) optimize the assay for buffer conditions, reagent concentrations, timing, stopping, order of addition, plate type and assay volume.

2020年9月1日 · Using these assays, the ability of test compounds to inhibit the kinase activity can be determined. How do kinase assays work? In general, the search for kinase inhibitors requires a very targeted approach to decrease the possibility of confounding effects.

A kinase assay is used to determine whether one protein is capable of phosphorylating another protein. As mentioned previously, protein kinases are enzymes that catalyze the covalent transfer of phosphate from adenosine triphosphate (ATP) to a substrate protein.

2018年12月10日 · A kinase assay works by simply measuring the activity of the kinase. A kinase is an enzyme that aids in the transfer of a phosphate from ATP to another specific molecule. They regulate many activities in the human body.

The classic continuous coupled-enzyme assay is the pyruvate kinase and lactate dehydrogenase system (Kornberg and Pricer, 1951). During the reaction, pyruvate kinase uses the ADP produced by the target enzyme to convert phosphoenolpyruvate to pyruvate and in …

This article will review some of the major kinase assay technologies on the market, with particular emphasis on the newest systems. We will describe the physical principles, the practical advantages and drawbacks, and the potential applications of these technologies in …

2006年8月3日 · The kinase assay procedure described here can be used to measure the activity of a purified protein kinase or to monitor its activity during purification from a cell extract.

2015年11月1日 · Promega's Kinase-Glo™ luminescent kinase assay detects kinase activity by measuring the depletion of ATP, making it similar to technologies that measure the production of ADP [37]. This method is a universal biochemical assay that can be used for any combination of kinase and substrate, regardless of the nature of the substrate such as ...

2019年12月19日 · We demonstrate proof of concept by designing an assay that jointly measures activity of five pleiotropic signaling kinases: Akt, IκB kinase (IKK), c-jun N-terminal kinase (JNK),...