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Fluorescent probe for detecting cell hydroxyl radical, and synthesis method and use

A fluorescent probe and free radical technology, applied in the field of biological detection technology and clinical medical detection, to achieve the effect of good permeability, good selectivity and short response time

Inactive Publication Date: 2007-08-22
SHANDONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fluorescent probes for intracellular nitric oxide and hydrogen peroxide detection and imaging have been reported (Sasaki E., Kojima H., Nishimatsu H., Urano Y., Kikuchi K., Hirata Y., Nagano T., J .Am.Chem.Soc.2005, 127, 3684; Xu K H., Tang B., Huang H., Yang G.W., Chen Z.Z., Li P., An L.G., Chem.Commun.2005, 48, 5974), However, fluorescent probes based on the principle of fluorescence resonance energy transfer for the detection and imaging of intracellular hydroxyl radicals have not been reported yet.

Method used

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  • Fluorescent probe for detecting cell hydroxyl radical, and synthesis method and use
  • Fluorescent probe for detecting cell hydroxyl radical, and synthesis method and use
  • Fluorescent probe for detecting cell hydroxyl radical, and synthesis method and use

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Probe Synthesis

[0039]

[0040] In the formula

[0041] AGGGTTAGGG: DNA oligonucleotide chain single strand, any base sequence, the number of bases is 2-30;

[0042] Fluorescein: X 1 , X 2 = F, Cl, CH 3 , OCH 3 ;

[0043] SH: Mercapto

[0044] Gold nanoparticles: 3-32nm in diameter.

[0045] a. Take 150.0 mL of chloroauric acid solution with a weight concentration of 0.01% in a 250 mL Erlenmeyer flask, heat to reflux, add 5.0 mL of trisodium citrate solution with a weight concentration of 2.0% under stirring, and continue to reflux for 20 minutes. placed under cooling to obtain a dark red gold nanoparticle solution; the transmission electron microscope results showed that the average particle diameter of the particles was 15.5nm, and the particle concentration was 1.2×10 according to the complete reaction estimation. 15 / L.

[0046] b. Mix the gold nanoparticle solution with the 5' end-modified fluorescein and the 3' end-modified thiol-modified DNA single ...

Embodiment 2

[0048] a. Take 150.0 mL of chloroauric acid solution with a weight concentration of 0.02% in a 250 mL conical flask, heat to reflux, add 7.0 mL of trisodium citrate solution with a weight concentration of 1.0% under stirring, and continue to reflux for 10 minutes. Place it under cooling to obtain a deep red gold nanoparticle solution; the transmission electron microscope results show that the average particle diameter of the particles is 15.9nm, and the particle concentration is 1.2×10 according to the complete reaction estimation. 15 / L.

[0049] b. Mix the gold nanoparticle solution with the 5'-end modified fluorescein and the 3'-end sulfhydryl-modified DNA single-strand according to the ratio of 1:500, leave it at room temperature for 12 hours, and then adjust the pH to 7.8 with phosphate buffer ;Then the sodium chloride solution that is 0.5mol / L is mixed with the sodium chloride solution whose concentration is 0.5mol / L through adjusting the pH value, and is placed at room ...

Embodiment 3

[0051] a. Take 150.0 mL of chloroauric acid solution with a weight concentration of 0.015% in a 250 mL Erlenmeyer flask, heat to reflux, add 6.0 mL of trisodium citrate solution with a weight concentration of 1.5% under stirring, and continue to reflux for 15 minutes. placed under cooling to obtain a dark red gold nanoparticle solution; the transmission electron microscope results showed that the average particle diameter of the particles was 15.7nm, and the particle concentration was 1.2×10 according to the complete reaction estimation. 15 / L.

[0052] b. Mix the gold nanoparticle solution with the 5'-end modified fluorescein and the 3'-end sulfhydryl-modified DNA single-strand according to the ratio of 1:350, leave it at room temperature for 18 hours, and then adjust the pH to 7.5 with phosphate buffer ; then the mixed solution through adjusting the pH value and the sodium chloride solution whose concentration is 3mol / L are mixed evenly by 1: 0.2 parts by volume, and placed ...

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Abstract

The invention provides a fluorescence probe, the synthetic method and the use which is to test the cell hydroxyl free radical. The process is: a. heat and chloroauric acid of 150.0 weight concentration and back flow, with mixing, the sodium citrate of 5.0-7.0 is added into the solution to back flow for 10-20min, then to cool in room temperature to get the golden nanometer particle solution; b. mix the particle with the DNA single chain with 5'end modified fluorescein and 3'end modified sulfydryl according to 1:200-500, then to set in room temperature for 1224h, last to adjust the pH by the phosphoric buffer; next to mix the mixture with the 0.5-5mol / L NaCl according to 1:0.1-0.3(V / V) which is set in room temperature for 35-45h; last to centrifugate to remove the upper solution and the deposition is solved in the 0.1-0.3mol / L phosphoric buffer which is the fluorescence probe solution.

Description

Technical field: [0001] The invention belongs to the fields of biological detection technology and clinical medical detection, and relates to a fluorescent probe for detecting cell hydroxyl free radicals; at the same time, it relates to a synthesis method of the fluorescent probe; in addition, it also relates to applications of the fluorescent probe. Background technique: [0002] Various free radicals are produced in the metabolic process of life activities, and the aging of organisms and the occurrence of many diseases are closely related to free radicals. Free radicals can mediate many important reaction processes of life activities, participate in cellular immune function, destructively modify the chemical structure of tissue cells, and damage the morphology and membrane function of normal tissue cells (Witz G., Proc.Soc.Exp.Biol Med. 1991, 198:675; Feig D.I., Reid T.M., Leob L.A., Cancer Res. 1994, 54:1890s; Finkl T., Holbrook N.J., Nature 2000, 408:239). Hydroxyl free...

Claims

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Application Information

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IPC IPC(8): G01N33/60G01N33/52G01N33/53
Inventor 唐波张宁徐克花禚林海陈蓁蓁
Owner SHANDONG NORMAL UNIV
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