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Method for detecting arginine kinase

A technology of arginine kinase and detection method, which is applied in the direction of measuring devices, material stimulation analysis, instruments, etc., can solve the problems of complex immune methods, time-consuming and labor-intensive, and achieve the effect of simple operation

Inactive Publication Date: 2017-09-19
XIAMEN UNIV
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Problems solved by technology

However, the immunization method is more complicated, time-consuming and labor-intensive.

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  • Method for detecting arginine kinase

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Embodiment Construction

[0017] The following embodiments will further illustrate the present invention in conjunction with the accompanying drawings.

[0018] The present invention comprises the following steps:

[0019] 1) Colloidal gold nanoparticles with a size of 30nm were synthesized by sodium citrate reduction method, and then 4-MBA was added overnight. Then remove free 4-MBA by centrifugation, and finally redisperse the precipitate in ultrapure water;

[0020] 2) Dissolve the substrate peptide in water so that the final concentration is 1mg / mL, take 2uL of the substrate peptide, and then add 1uLATP and MgCl respectively 2 , and finally add different concentrations of arginine kinase to the substrate peptide, the final volume is 100uL, and place 100uL of the reaction solution in a 40°C water bath for 3h;

[0021] 3) Take 5uL of the above-mentioned reaction solution and add it to 200uL of gold nanoparticles, mix well and perform Raman spectrum test, and finally obtain a standard curve.

[002...

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Abstract

The invention provides a method for detecting arginine kinase, and relates to arginine kinase. The method includes synthesizing colloidal nano-gold by the aid of sodium citrate reduction processes, adding 4-MBA (4-maleimidobutyric acid) into the colloidal nano-gold, then removing the free 4-MBA by means of centrifugation and re-dispersing precipitates in ultra-pure water; dissolving substrate peptides in water, adding ATP (adenosine triphosphate) and MgCl2 into the substrate peptides, adding the arginine kinase into the substrate peptides to obtain reaction liquid, placing the reaction liquid in a water bath kettle and keeping the temperature of the reaction liquid constant; adding the reaction liquid into the nano-gold, uniformly mixing the reaction liquid and the nano-gold with each other and testing Raman spectra to ultimately obtain standard curves. The method has the advantages that intracellular signal transduction, protein structures and the like can be regulated by kinase, and accordingly the method for detecting the arginine kinase is quite important for detecting the kinase.

Description

technical field [0001] The invention relates to arginine kinase, in particular to a method for detecting arginine kinase (McsB). Background technique [0002] Due to the high bond energy of the phosphorus-nitrogen bond and its easy hydrolysis under acidic conditions, the detection of arginine kinase is facing great challenges. Since the first arginine kinase was discovered in 2009, many works have been devoted to the detection of kinases. At present, the main method is immunodetection: using phosphorylated arginine to synthesize antibodies, and then using antibodies to detect arginine Kinases (see: Ouyang, H., Fu, C., Fu, S. Org Biomol Chem, 2016, 14, 6, 1925). However, the method of immunization is more complicated, time-consuming and labor-intensive. Therefore, there is a need for a simple and rapid kinase detection method with few operating steps. Contents of the invention [0003] The purpose of the present invention is to provide a detection method of arginine kina...

Claims

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Application Information

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IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 赵玉芬蔡华欢黄碧玲林荣灿许鹏翔刘艳
Owner XIAMEN UNIV
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