Visualization of Kinase Activity with FRET‐Based Activity Biosensors
Genetically encodable FRET-based kinase activity reporters (KARs) enable real-time monitoring of kinase activity dynamics in living cells with high spatiotemporal resolution. This unit describes a general protocol for utilizing KARs to visualize kinase activity in living mammalian cells with fluorescence microscopy. Curr. Protoc. Mol. Biol. 91:18.15.1-18.15.9. © 2010 by John Wiley & Sons, Inc.
Keywords: kinase activity reporter; FRET; live-cell imaging
Basic Protocol: Measuring Protein Kinase Activity Using Genetically Encoded Reporters
Figure 18.15.2 Diffusible (left) and plasma membrane-targeted (right) AKAR.
Figure 18.15.3 Excitation and emission spectra for CFP and YFP.
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This paper provides detailed information on how to design and characterize FRET-based kinase activity biosensors.
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This paper provides information on the design of genetically encodable, FRET-based kinase activity biosensors and several examples of their use in studying kinase regulation in living cells with high spatiotemporal resolution.