Time-resolved fluorescence resonance energy transfer assay

Newman, M. and Josiah, S., Utilization of fluorescence polarization and time resolved fluorescence resonance energy transfer assay formats for SAR studies SRC kinase as a model system, /. Biomol. Screen., 9, 525, 2004. 

Ozers, M.S., Marks, B.D., Gowda, K., Kupcho, K.R., Ervin, K.M., De Rosier, T., Qadir, N., Eliason, H.C., Riddle, S.M. and Shekhani, M.S. (2007) The androgen receptor T877A mutant recruits LXXLL and FXXLF peptides differently than wild-type androgen receptor in a time-resolved fluorescence resonance energy transfer assay. Biochemistry, 46, 683-695. 

Schroter, T. et al. 2008. Comparison of miniaturized time-resolved fluorescence resonance energy transfer and enzyme-coupled luciferase high-throughput screening assays to discover inhibitors of Rho-kinase II (ROCK-II). J. Biomol. Screen. 13, 17-28. 

FIGURE 4.1 Assays commonly used in GPCR research. SPA = scintillation proximity assay FP = fluorescence polarization TR-FRET = time-resolved fluorescence resonance energy transfer FCS = fluorescence correlation spectroscopy SeAP = secreted alkaline phosphate TF = transcription factor EFC = enzyme fragment complementation DMR = dynamic mass redistribution CDS = cellular dielectric spectroscopy. 

Zhou, V. et al. 2004. A time-resolved fluorescence resonance energy transfer-based HTS assay and a surface plasmon resonance-based binding assay for heat shock protein 90 inhibitors. Anal. Biochem. 331, 349-357. 

Another example of improved sensitivity due to modulation of lanthanide photophysics by ancillary ligands can be found in the europium and terbiiun chelates used in time-resolved fluorescence resonance energy transfer (TR-FRET) immunoassays (100,101). Due to their line-type emissions and long decay times, the lanthanide chelate is used as a donor, with some visible-absorbing dye such as Alexa 647 or a rhodamine derivative as the acceptor. Without the helper ligand, the lanthanides would be unprotected from solvent and have much shorter decay times, making them unsuitable for such an assay. 

Laitala V, Hemmila L. Homogeneous assay based on low quan- 83. turn yield Sm(lll)-donor and anti-Stokes shift time-resolved fluorescence resonance energy-transfer measurement. Analyt. Chim. 

Time- and frequency-domain fluorescence methods have been used in different fields for several purposes, such as distinguishing sample constituents whose fluorescence spectra overlap one another, to distinguish the fluorescence of an analyte from background scattering or luminescence of other sample constituents, in combination with fluorescence resonance energy transfer (FRET) assays to combine the benefits of FRET and time-resolved fluorescence. A listing of some selected applications is given in Table 3. 

Stenroos K, Hurskainen P, Eriksson S et al (1998) Homogeneous time-resolved 1L-2-IL-2R alpha assay using fluorescence resonance energy transfer. Cytokine 10 495 99... 

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