A rapid SNAP-tag fluorogenic probe based on an environment-sensitive fluorophore for no-wash live cell imaging

ACS Chem Biol. 2014 Oct 17;9(10):2359-65. doi: 10.1021/cb500502n. Epub 2014 Aug 14.

Abstract

One major limitation of labeling proteins with synthetic fluorophores is the high fluorescence background, which necessitates extensive washing steps to remove unreacted fluorophores. In this paper, we describe a novel fluorogenic probe based on an environment-sensitive fluorophore for labeling with SNAP-tag proteins. The probe exhibits dramatic fluorescence turn-on of 280-fold upon being labeled to SNAP-tag. The major advantages of our fluorogenic probe are the dramatic fluorescence turn-on, ease of synthesis, high selectivity, and rapid labeling with SNAP-tag. No-wash labeling of both intracellular and cell surface proteins was successfully achieved in living cells, and the localization of these proteins was specifically visualized.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cytosol / metabolism*
  • Flow Cytometry
  • Fluorescent Dyes / chemistry*
  • HCT116 Cells
  • Humans
  • Molecular Imaging / methods*
  • Molecular Probes*
  • Proteins / metabolism*

Substances

  • Fluorescent Dyes
  • Molecular Probes
  • Proteins