A Rapid SNAP-Tag Fluorogenic Probe Based on an Environment-Sensitive Fluorophore for No-Wash Live Cell Imaging
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- 作者：Tao-Kai Liu ; 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
- 刊名：ACS Chemical Biology
- 出版年：2014
- 出版时间：October 17, 2014
- 年：2014
- 卷：9
- 期：10
- 页码：2359-2365
- 全文大小：408K
- ISSN：1554-8937
文摘
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.