Substrates for Improved Live-Cell Fluorescence Labeling of SNAP-tag

Author(s): Ivan R. Correa, Brenda Baker, Aihua Zhang, Luo Sun, Christopher R. Provost, Grazvydas Lukinavicius, Luc Reymond, Kai Johnsson, Ming-Qun Xu

Journal Name: Current Pharmaceutical Design

Volume 19 , Issue 30 , 2013

Become EABM
Become Reviewer
Call for Editor


The SNAP-tag labeling technology provides a simple, robust, and versatile approach to the imaging of fusion proteins for a wide range of experimental applications. Owing to the specific and covalent nature of the labeling reaction, SNAP-tag is well suited for the analysis and quantification of fused target protein using fluorescence microscopy techniques. In this report, we present our most recent findings on the labeling of SNAP-tag fusion proteins both in vitro and in cell culture with SNAP-tag substrates derived from single regioisomers of carboxyrhodamine dyes. Carboxyrhodamines are invaluable fluorescent dyes for biotechnology applications including DNA sequencing, detection on microarrays, and fluorescence in situ hybridization. We found that SNAP-tag reacts preferentially with the 6-positional regioisomer of carboxyrhodamine fluorescent dyes, whereas the 5-regioisomer predominantly contributes to background fluorescence. Our experimental study also indicates that benzylchloropyrimidine (CP) conjugates of 6-carboxyrhodamines exhibit a dramatic increase in the signal-to-noise ratio of fluorescently labeled cellular proteins compared to the benzylguanine (BG) conjugates, presumably due to higher cell permeability. These new SNAP-tag substrates based on pure 6-regioisomers can significantly improve fluorescence labeling in live cells and should become powerful tools for bioimaging applications.

Keywords: Cell imaging, covalent labeling, fluorescent probes, protein modifications.

Rights & PermissionsPrintExport Cite as

Article Details

Year: 2013
Page: [5414 - 5420]
Pages: 7
DOI: 10.2174/1381612811319300011
Price: $65

Article Metrics

PDF: 57