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

TK Liu, PY Hsieh, YD Zhuang, CY Hsia… - ACS Chemical …, 2014 - ACS Publications
TK Liu, PY Hsieh, YD Zhuang, CY Hsia, CL Huang, HP Lai, HS Lin, IC Chen, HY Hsu
ACS Chemical Biology, 2014ACS Publications
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 …
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.
ACS Publications