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.
Tao-Kai Liu  1 ,  Pei-Ying Hsieh ,  Yu-De Zhuang ,  Chi-Yang Hsia ,  Chi-Ling Huang ,  Hsiu-Ping Lai ,  Hung-Sheung Lin ,  I-Chia Chen ,  Hsin-Yun Hsu ,  Kui-Thong Tan
Affiliations
  • 1. Department of Chemistry and ‡Frontier Research Center on Fundamental and Applied Sciences of Matters, National Tsing Hua University , 101 Sec. 2, Kuang-Fu Rd., Hsinchu 30013, Taiwan (ROC).
PMID: 25105835 DOI: 10.1021/cb500502n
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.

Products