A kind of wash-free snap-tag probe excited by 488nm and preparation method thereof

A technology of probes and synthesis methods, applied in the field of protein labeling, can solve the problems of inability to obtain imaging results, increase of false signals, and inability to control the number and position, and achieve the effects of maintaining accuracy, low-cost synthetic materials, and simple and general methods.
CN111333618BActive Publication Date: 2022-03-18DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
Publication Date
2022-03-18

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Abstract

The invention provides a wash-free SNAP-tag probe excited at 488nm and a preparation method thereof. The wash-free SNAP-tag probe introduces a spiro ring rigid structure into a disubstituted naphthalimide dye at the 4,5-position And design and synthesize a series of wash-free SNAP-tag probes that can be used for 488nm excitation, and its structural formula is shown in (1). The introduction of the rigid structure not only limits the intramolecular torsion, improves the stability and brightness of the fluorescent molecules, but also increases the planarity of the probe molecules. This causes the SNAP‑tag probe molecules to form a strong π‑π interaction in water, resulting in a quenching of the fluorescence. After combining with SNAP-tag, the fluorescence of the probe molecule is released, which can be enhanced up to 28 times. This series of probes can achieve specific labeling of SNAP-tag in living cells and achieve wash-free fluorescence imaging. In addition, due to the improvement of stability and brightness, this series of probes can also be used in SIM (Structured Light Illumination Microscopy), STED (Stimulated Emission Depletion) and other super-resolution fluorescence imaging fields.
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Description

technical field

[0001] The invention belongs to the field of protein labeling, and in particular relates to a 488nm-excited wash-free SNAP-tag probe and a preparation method thereof. Background technique

[0002] Fluorescence imaging technology has gradually become a powerful tool for studying protein function from the cellular level to the individual level. Due to the advantages of small size, broad fluorescence emission spectrum, and a variety of fluorescent colors, organic small molecule fluorescent dyes have gradually become a substitute for fluorescent proteins in the field of protein labeling. However, organic small molecule dyes are exogenous species, and the problem is that they cannot be derived from cells like fluorescent proteins, so the number and position in cells cannot be controlled. In order to solve this problem, chemists have developed a variety of bioorthogonal methods to covalently attach small molecule dyes to target proteins, so that the location and f...

Claims

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