A method to quantify FRET stoichiometry with phasor plot analysis and acceptor lifetime ingrowth.
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ABSTRACT:
SUBMITTER: Chen W
PROVIDER: S-EPMC4375440 | biostudies-literature | 2015 Mar
REPOSITORIES: biostudies-literature
Publications
A method to quantify FRET stoichiometry with phasor plot analysis and acceptor lifetime ingrowth.
Chen WeiYue W Avezov Edward E Schlachter Simon C SC Gielen Fabrice F Laine Romain F RF Harding Heather P HP Hollfelder Florian F Ron David D Kaminski Clemens F CF
Biophysical journal 20150301 5
FRET is widely used for the study of protein-protein interactions in biological samples. However, it is difficult to quantify both the FRET efficiency (E) and the affinity (Kd) of the molecular interaction from intermolecular FRET signals in samples of unknown stoichiometry. Here, we present a method for the simultaneous quantification of the complete set of interaction parameters, including fractions of bound donors and acceptors, local protein concentrations, and dissociation constants, in eac ...[more]
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