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Fluorescent Labeling of SNAP-Tagged Proteins in Cells

  • Gražvydas Lukinavičius
  • Luc Reymond
  • Kai JohnssonEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1266)

Abstract

One of the most prominent self-labeling tags is SNAP-tag. It is an in vitro evolution product of the human DNA repair protein O 6-alkylguanine-DNA alkyltransferase (hAGT) that reacts specifically with benzylguanine (BG) and benzylchloropyrimidine (CP) derivatives, leading to covalent labeling of SNAP-tag with a synthetic probe (Gronemeyer et al., Protein Eng Des Sel 19:309–316, 2006; Curr Opin Biotechnol 16:453–458, 2005; Keppler et al., Nat Biotechnol 21:86–89, 2003; Proc Natl Acad Sci U S A 101:9955–9959, 2004). SNAP-tag is well suited for the analysis and quantification of fused target protein using fluorescence microscopy techniques. It provides a simple, robust, and versatile approach to the imaging of fusion proteins under a wide range of experimental conditions.

Key words

Snap-tag Synthetic fluorophores Living and fixed cells Covalent labeling Self-labeling tags Fluorescence microscopy Episomal protein expression 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Gražvydas Lukinavičius
    • 1
  • Luc Reymond
    • 1
  • Kai Johnsson
    • 1
    Email author
  1. 1.Institute of Chemical Sciences and Engineering, NCCR Chemical BiologyÉcole Polytechnique Fédérale de Lausanne (EPFL)LausanneSwitzerland

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