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Preparation method of nano fluorescent probe for detecting deubiquitination enzyme

A nano-fluorescent probe and deubiquitinating enzyme technology, which is applied in the field of preparation of nano-fluorescent probes, can solve the problems of easy extinguishment of fluorescence, achieve good sensitivity and selectivity, good selectivity, and short preparation cycle

Active Publication Date: 2020-05-15
SHANDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the fluorescence of lanthanide probes is easily extinguished in water and biosolvents due to competitive coordination

Method used

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  • Preparation method of nano fluorescent probe for detecting deubiquitination enzyme
  • Preparation method of nano fluorescent probe for detecting deubiquitination enzyme
  • Preparation method of nano fluorescent probe for detecting deubiquitination enzyme

Examples

Experimental program
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Effect test

Embodiment 1

[0057] (1) 1.02mmol 1,10-phenanthroline-5-amine and 1.7mL 3-(triethoxysilyl)propyl isocyanate were stirred in chloroform for 30min, and the chloroform was removed under vacuum at 25°C. The remaining yellow viscous mixture was stirred at 80 °C for another 20 hours. After the reaction, cold hexane was added to the mixture to obtain a pale yellow precipitate, which was then dissolved in methanol and filtered. Methanol was removed by evaporation and washed with cold hexane. The product phenanthroline-silicon was reprecipitated with alkane, and the prepared phenanthroline-silicon was dried in vacuum at 60°C.

[0058] (2) Disperse 500 mg of mesoporous silicon nanoparticles in 10 mL of dichloromethane solution containing 2 mg of phenanthroline-silicon. The reaction was carried out under nitrogen protection at 90 ° C for 12 hours. After the reaction, the prepared mesoporous silicon was collected by filtration. Porous silica-phenanthroline nanoparticles, rinsed with ethanol and dried u...

Embodiment 2

[0063] (1) 1.02mmol 1,10-phenanthroline-5-amine and 1.7mL 3-(triethoxysilyl)propyl isocyanate were stirred in chloroform for 30min, and the chloroform was removed under vacuum at 25°C. The remaining yellow viscous mixture was stirred at 80 °C for another 20 hours. After the reaction, cold hexane was added to the mixture to obtain a pale yellow precipitate, which was then dissolved in methanol and filtered. Methanol was removed by evaporation and washed with cold hexane. The product phenanthroline-silicon was reprecipitated with alkane, and the prepared phenanthroline-silicon was dried in vacuum at 60°C.

[0064] (2) Disperse 500 mg of mesoporous silicon nanoparticles in 10 mL of dichloromethane solution containing 8 mg of phenanthroline-silicon. The reaction was carried out under nitrogen protection at 90 ° C for 12 hours. After the reaction, the prepared mesoporous silicon was collected by filtration. Porous silica-phenanthroline nanoparticles, rinsed with ethanol and dried u...

Embodiment 3

[0069] (1) 1.02mmol 1,10-phenanthroline-5-amine and 1.7mL 3-(triethoxysilyl)propyl isocyanate were stirred in chloroform for 30min, and the chloroform was removed under vacuum at 25°C. The remaining yellow viscous mixture was stirred at 80 °C for another 20 hours. After the reaction, cold hexane was added to the mixture to obtain a pale yellow precipitate, which was then dissolved in methanol and filtered. Methanol was removed by evaporation and washed with cold hexane. The product phenanthroline-silicon was reprecipitated with alkane, and the prepared phenanthroline-silicon was dried in vacuum at 60°C.

[0070] (2) Disperse 500 mg of mesoporous silicon nanoparticles in 10 mL of dichloromethane solution containing 10 mg of phenanthroline-silicon. The reaction was carried out under nitrogen protection at 90° C. and stirred for 12 hours. After the reaction, the prepared mesoporous silicon was collected by filtration. Porous silica-phenanthroline nanoparticles, rinsed with ethano...

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Abstract

The invention belongs to the technical field of deubiquitination enzyme fluorescence detection, and particularly relates to a preparation method of a nano fluorescent probe for detecting deubiquitination enzyme. The preparation method comprises following steps: dispersing mesoporous silicon nano particles into a dichloromethane solution containing phenanthroline-silicon, carrying out reactions toobtain mesoporous silicon-phenanthroline nano particles; mixing mesoporous silicon-phenanthroline nano particles and TbCl3.6H2O in ethanol, refluxing, adding 1,3-diphenyl-1,3-propanedione, carrying out reactions to obtain a nano fluorescent probe precursor; suspending the nano fluorescent probe precursor in a polyethyleneimine water solution to coat polyethyleneimine to obtain a polyethyleneiminemodified nano fluorescent probe precursor; dispersing the polyethyleneimine modified nano fluorescent probe precursor into the mixed solution, and stirring to obtain the nano fluorescent probe. The preparation process is simple, the period is short; and the prepared nano fluorescent probe has good sensitivity and selectivity for detection of UCH-L1.

Description

technical field [0001] The invention belongs to the technical field of fluorescence detection of deubiquitinating enzymes, and in particular relates to a preparation method of a nanometer fluorescent probe used for detecting deubiquitinating enzymes. Background technique [0002] Ubiquitin is a small protein consisting of 76 amino acids. Ubiquitination is a process in which ubiquitin specifically modifies target proteins under the action of a series of specific enzymes, and regulates protein activity. During ubiquitination, it binds to the target protein by forming an isopeptide bond between the C-terminus of ubiquitin and the endonuclease of the target protein. Deubiquitinating enzymes can catalyze the hydrolysis of peptide bonds, thereby regulating the ubiquitination state of target proteins. So ubiquitination can be reversed by the action of deubiquitinating enzymes. A large class of deubiquitinating enzymes regulates this cellular process and plays an important role i...

Claims

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Application Information

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IPC IPC(8): C09K11/02C09K11/06G01N21/64
CPCC09K11/025C09K11/06G01N21/6428C09K2211/182C09K2211/1044
Inventor 王粤博梁妍妍王怀松崔辉艾兵
Owner SHANDONG UNIV OF TECH
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