| Assay Method Information | |
| | AKT IMAP Fluorescence Polarization Assay |
| Description: | A bead-based FP assay methodology, termed IMAP, has been developed for the serine/threonine kinase, AKT that allows for direct measurement of product formation. The assay design utilizes a fluoresceinated peptide substrate that, when phosphorylated by the kinase, binds to nanoparticles derivatized with trivalent metal cations through a metal-phospholigand interaction. The result of this bound fluoresceinated phosphorylated product is an increase in polarization signal caused by a decrease in the molecular mobility of the bound product. Reported values are an average of three independent binding curves utilizing an IMAP Akt Assay Kit (Molecular Devices). Reactions were conducted in wells with fluoresceinated peptide, ATP, and Akt1, and inhibitor in reaction buffer. The reactions were equilibrated for 1 h at room temperature then data points were collected and analyzed. |
| Affinity data for this assay | |
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