Technical Reference #683

683.

Estimating Protein-Protein Interaction Affinity In Living Cells Using Quantitative Forster Resonance Energy Transfer Measurements Huanmian Chen; Henry L. Puhl III; Stephen R. Ikeda, National Institute of Health, Journal of Biomedical Optics, 12(683), (2007)
Link To Paper

Abstract:
Abstract. We have previously demonstrated that Förster resonance energy transfer FRET efficiency and the relative concentration of donorand acceptor fluorophores can be determined in living cells usingthree-cube wide-field fluorescence microscopy.

Keywords:
Förster resonance energy transfer; protein-protein interaction; fluorescent proteins.

Materials & Methods:
HeLa cells ATCC Manassas Virginia suspended in minimal essential medium containing 10% bovine calf serum were plated onto glass-bottom tissue culture dishes MatTek Ashland Massachusetts and stored in a humidified atmosphere containing 5% CO2 in ai

Microscopic Technique
Fluorescence Microscopy

Cell Type(s)
HeLa