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A method for detecting overexpression of cell membrane proteins based on nano-fluorescent probes and its application

A nano fluorescent probe and cell membrane technology, applied in nano optics, nanotechnology, nanotechnology, etc., can solve the problems of high cost and dependence on large instruments, and achieve the effect of easy operation, easy promotion and stable effect

Active Publication Date: 2022-07-19
LIAONING UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above detection methods have certain limitations and problems, such as dependence on large instruments, high cost, timeliness, etc.

Method used

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  • A method for detecting overexpression of cell membrane proteins based on nano-fluorescent probes and its application
  • A method for detecting overexpression of cell membrane proteins based on nano-fluorescent probes and its application
  • A method for detecting overexpression of cell membrane proteins based on nano-fluorescent probes and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A method for detecting the overexpression of cell membrane proteins based on nano-fluorescent probes, comprising the following steps:

[0029] (1) Preparation of nano-fluorescent probes

[0030] Weigh 2g (13mmol) histidine and 0.024g valine (2mmol), dissolve in 20mL ultrapure water, add 0.18g (3mmol) ethylenediamine, after complete ultrasonic dissolution, under the condition of 210 ℃ in the reactor After heating for 15h, nano-fluorescent probes were obtained.

[0031] figure 1 is the scanning electron microscope image of the prepared nano-fluorescent probe, from figure 1 It can be seen that the average particle size of the prepared nano-fluorescent probes is about 4 nm, and the distribution is uniform, forming uniformly distributed spherical particles.

[0032] Figure 2A It is the prepared nano-fluorescent probe sample, and the fluorescence spectrum is obtained by fluorescence detection at the optimal excitation wavelength with a fluorescence spectrophotometer. De...

Embodiment 2

[0040] Screening methods for drugs that inhibit the overexpression of cell membrane proteins are as follows:

[0041] 1. Use a cell scraper to separate the overexpressing microglia from the culture flask, and seed them in 26 6-well plates to make the number of cells 5×10 5 indivual. After 4 hours of cell adhesion, 23 kinds of drugs with a concentration of 5 μg / ml, such as sulindac and metformin hydrochloride, were added, and placed in 5% CO. 2 Incubate for 24h in incubator. After taking out the cells and washing with cold PBS three times, the cells were separated from the 6-well plate with a cell scraper, then centrifuged at 1000 r / min at 4 °C for 4 min, and redispersed in 1 ml of 10 mmol / LTris-HCl buffer solution. Under the condition of 300w, the cell disruptor was sonicated for 3 s, with an interval of 7 s, and the cells were disrupted for 3 min. According to the differential centrifugation method, the separation of cell membrane proteins was carried out in a low temperat...

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Abstract

The invention relates to a method for detecting the overexpression of cell membrane proteins based on nano-fluorescent probes and its application. In a 96-well plate, add the cell membrane protein solution and the nano-fluorescent probe solution, incubate at 20-25°C for 5-10min, and then detect the fluorescence intensity. The preparation method of the nano-fluorescent probe solution is as follows: completely dissolving the carbon source with water, adding small molecule ligands to the solution for doping, and after ultrasonic dissolving, heating and reacting at 190-220° C. to obtain the nano-fluorescent probe, The nano-fluorescent probe was filtered, dialyzed, lyophilized and then dispersed in a PBS buffer solution to obtain a nano-fluorescent probe solution. The method of the invention can identify cell membrane proteins in different response states, and also has the characteristics of good biocompatibility, safety and non-toxicity, stable effect and the like. The overexpression of the cell membrane protein can be specifically recognized, and the preparation method is simple in process, easy to operate, low in preparation cost and easy to popularize.

Description

technical field [0001] The invention belongs to the technical field of fluorescent nanomaterials, and in particular relates to a method for detecting the overexpression of cell membrane proteins based on nano fluorescent probes and its application. Background technique [0002] Protein is the material basis of life, organic macromolecules, the basic organic matter that constitutes cells, and the main undertaker of life activities. There is no life without protein. Proteins are involved in every cell and all vital components in the body. Protein is an important part of body cells and the main raw material for human tissue renewal and repair. It mainly maintains the normal metabolism of the body and the transport of various substances in the body. For example: carrier protein is essential to maintain the normal life activities of the human body. Various substances can be carried in the body. For example, hemoglobin transports oxygen (the red blood cell renewal rate is 2.5...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64C09K11/06B82Y40/00B82Y20/00
CPCG01N21/6428C09K11/06B82Y20/00B82Y40/00
Inventor 刘学毕秀丽刘禹霖陈扬
Owner LIAONING UNIVERSITY