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Azide-specific labeling of biomolecules by Staudinger-Bertozzi ligation phosphine derivatives of fluorescent probes suitable for single-molecule fluorescence spectroscopy.

Methods in enzymology (2010-06-29)
Anirban Chakraborty, Dongye Wang, Yon W Ebright, Richard H Ebright
ABSTRACT

We describe the synthesis of phosphine derivatives of three fluorescent probes that have a brightness and photostability suitable for single-molecule fluorescence spectroscopy and microscopy: Alexa488, Cy3B, and Alexa647. In addition, we describe procedures for use of these reagents in azide-specific, bioorthogonal labeling through Staudinger-Bertozzi ligation, as well as procedures for the quantitation of labeling specificity and labeling efficiency. The reagents and procedures of this report enable chemoselective, site-selective labeling of azide-containing biomolecules for single-molecule fluorescence spectroscopy and microscopy.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
8-(6-Aminohexyl)aminoadenosine 3′:5′-cyclic monophosphate, ~98%