Nature Methods
- 5, 401 - 403 (2008)
Published online: 20 April 2008; | doi:10.1038/nmeth.1207
Fluorescent protein FRET pairs for ratiometric imaging of dual biosensorsHui-wang Ai1, Kristin L Hazelwood2, Michael W Davidson2 & Robert E Campbell11
University of Alberta, Department of Chemistry, 11227 Saskatchewan Drive, Edmonton, Alberta T6G 2G2, Canada. 2
National High Magnetic Field Laboratory and Department of Biological Science, Florida State University, 1800 East Paul Dirac Drive, Tallahassee, Florida 32310, USA.
Correspondence should be addressed to Robert E Campbell robert.e.campbell@ualberta.ca Fluorescence resonance energy transfer (FRET) with fluorescent proteins is a powerful method for detection of protein-protein interactions, enzyme activities and small molecules in the intracellular milieu. Aided by a new violet-excitable yellow-fluorescing variant of Aequorea victoria GFP, we developed dual FRET–based caspase-3 biosensors. Owing to their distinct excitation profiles, each FRET biosensor can be ratiometrically imaged in the presence of the other.
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