A Rapid SNAP-Tag Fluorogenic Probe Based on an Environment-Sensitive Fluorophore for No-Wash Live Cell Imaging
A Rapid SNAP-Tag Fluorogenic Probe Based on an Environment-Sensitive Fluorophore for No-Wash Live Cell Imaging
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DOI：
10.1021/cb500502n
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发表时间：
2014-10-01
影响因子：
4
通讯作者：
Tan, Kui-Thong
中科院分区：
文献类型：
--
作者：
Liu, Tao-Kai;Hsieh, Pei-Ying;Tan, Kui-Thong
用合成荧光团标记蛋白质的一个主要限制是高荧光背景，这需要大量的洗涤步骤以去除未反应的荧光团。在本文中，我们描述了一种基于环境敏感荧光团的新型荧光探针，用于标记 SNAP 标签蛋白。该探针在标记 SNAP 标签后，荧光强度提高了 280 倍。我们的荧光探针的主要优点是显着的荧光开启、易于合成、高选择性以及使用 SNAP 标签快速标记。在活细胞中成功实现了细胞内和细胞表面蛋白质的免洗标记，并且这些蛋白质的定位被特异性地可视化。
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.