Method of determining energy transfer efficiency of a donor- acceptor pair of fluorescent dyes

摘要

The energy transfer efficiency of a donor-acceptor pair of fluorescent dyes can be determined by first measuring the fluorescence and absorption spectra of donor-protein, acceptor-protein, mixture of donor-protein and acceptor-protein, and donor-acceptor-protein conjugates, then separating the respective spectra into their respective donor- protein complex and acceptor-protein complex components using multiple linear regression, and then determining the transfer efficiency on the basis of the quenching of the donor fluorescence based on the spectral data thus obtained.