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Citation: QIAO Qinglong, ZHOU Wei, CHEN Jie, LIU Wenjuan, MIAO Lu, YIN Wenting, XU Zhaochao. Combined application of SNAP-tag and fluorescence technique in in-situ protein analysis[J]. Chinese Journal of Chromatography, ;2019, 37(8): 872-877. doi: 10.3724/SP.J.1123.2019.03020 shu

Combined application of SNAP-tag and fluorescence technique in in-situ protein analysis

Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China

Received Date: 19 March 2019

Fund Project: National Natural Science Foundation of China (No. 21878286).

The visualization of the microcosmic behavior of proteins in vivo is the key to real-time monitoring of proteins. A series of wash-free SNAP-tag probes were designed and synthesized based on the combination of SNAP-tag and small organic molecule fluorescent dyes. SNAP-tag, which specifically recognized O6-benzylguanine, could be labeled with a fluorophore (e. g., 1,8-naphthalimide) through the formation of covalent bonds. Furthermore, the change from a hydrophilic environment to the hydrophobic cavum of SNAP-tag realized a 2-13-fold enhancement in fluorescence. Through the fusion of SNAP-tag and the target protein, the probes could recognize the mitochondrial proteins (e. g., cytochrome oxidase, Cox8A) and nuclear proteins (e. g., H2B) in living cells. Besides, the fluorescent probes allowed the in-situ real-time monitoring of proteins without washing.
- SNAP-tag,
- in-situ,
- organic small molecule fluorescent dyes,
- wash-free fluorescent probes
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