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Curr Protoc Chem Biol. 2011;3(4):153-162. Epub 2011 Dec 1.

Copper-Catalyzed Azide-Alkyne Click Chemistry for Bioconjugation.

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  • 1Department of Chemistry, The Scripps Research Institute, La Jolla, California.

Abstract

The copper-catalyzed azide-alkyne cycloaddition reaction is widely used for the connection of molecular entities of all sizes. A protocol is provided here for the process with biomolecules. Ascorbate is used as reducing agent to maintain the required cuprous oxidation state. Since these convenient conditions produce reactive oxygen species, five equivalents of a copper-binding ligand is used with respect to metal. The ligand both accelerates the reaction and serves as a sacrificial reductant, protecting the biomolecules from oxidation. A procedure is also described for testing the efficiency of the reaction under desired conditions for purposes of optimization, before expensive biological reagents are used.

PMID:
22844652
[PubMed]
PMCID:
PMC3404492
Free PMC Article
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