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Published October 18, 2010 | Accepted Version + Supplemental Material
Journal Article Open

Live-Cell Imaging of Cellular Proteins by a Strain-Promoted Azide–Alkyne Cycloaddition

Abstract

Live and let dye: Three coumarin-cyclooctyne conjugates have been used to label proteins tagged with azidohomoalanine in Rat-1 fibroblasts. All three fluorophores labeled intracellular proteins with fluorescence enhancements ranging from eight- to 20-fold. These conjugates are powerful tools for visualizing biomolecule dynamics in living cells.

Additional Information

© 2010 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. Article first published online: 10 Sep. 2010. We thank Rebecca E. Connor, Julie C. Liu, Marissa L. Mock, Tae Hyeon Yoo, Jay A. Codelli, and Brian C. Dickinson for advice and assistance. We thank Scott E. Fraser, Chris Waters, and the Biological Imaging Center of the Beckman Institute at Caltech for advice on microscopy, Mona Shahgholi for assistance with mass spectrometry, and Rochelle Diamond and Diana Perez for assistance with flow cytometry. We thank Mandy K. S. Vink for Aha and Stacey A. Maskarinec for cell lines. This work was supported by NIH grant GM62523 to D.A.T. and GM66047 to C.R.B., by the ARO Institute for Collaborative Biotechnologies, by a Hertz Foundation Fellowship and PEO Scholar Award to K.E.B. , by NIH postdoctoral fellowships to J.D.F. and B.P.S., an NSF and NDSEG fellowship to J.M.B., and an NDSEG fellowship to M.J.H.

Attached Files

Accepted Version - nihms273547.pdf

Supplemental Material - cbic_201000419_sm_miscellaneous_information.pdf

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