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Method for determining cortex actin by electrochemical luminescence

An actin and electrochemical technology, which is applied in the field of analytical chemistry and electrochemiluminescence detection, can solve the problems of complex operation and expensive equipment, and achieve the effect of low material cost, low cost and high sensitivity

Active Publication Date: 2013-12-11
石家庄卢米特生物科技有限公司
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Problems solved by technology

These methods have their own advantages, and can meet the detection requirements of cortical actin to varying degrees, but the method is complicated to operate and the equipment is expensive.

Method used

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  • Method for determining cortex actin by electrochemical luminescence
  • Method for determining cortex actin by electrochemical luminescence
  • Method for determining cortex actin by electrochemical luminescence

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preparation example Construction

[0032] The preparation of electrochemiluminescent probe is characterized in that it comprises the following steps:

[0033] Preferably, 2.0 mg of the peptide is dissolved in 10 ml of absolute ethanol, and then 5 mg of Ru-NHS (Ru(bpy) 2 (dcbpy)NHS) solution, stirred and incubated overnight at 4°C to obtain an electrochemiluminescent probe (Ru-peptide).

[0034] The preparation of electrochemiluminescent probe sensor is characterized in that it comprises the following steps:

[0035] Preferably, gold nanoparticles and graphene-modified electrodes are soaked in 50 μL of electrochemiluminescence probe solution (phosphate buffer solution pH 7.4 + 5 mM TCEP (trichloroethyl phosphate)), and incubated overnight at 4 °C with stirring, washing. Then soak in 100 μL 1.0×10 -3 M MCH reacted for half an hour, then washed the electrode three times with pH 7.4 phosphate buffer solution, dried the surface of the electrode, and obtained. Store at 4 °C for later use.

[0036] Electroch...

Embodiment

[0043] Example: A method for measuring cortical actin by electrochemiluminescence

[0044] 1. Experimental part

[0045] 1.1 Instruments and reagents

[0046] Carboxylated ruthenium pyridine (Ru(bpy) 2 (dcbpy)NHS: bis(2,2'-bipyridine) (2,2'-pyridine-4,4'-dicarbonate) ruthenium succinamide, Ru-NHS), N,N'-dicyclohexylcarbodi Imine (EDC), thermolysin, TPA (Sigma, USA); cortical actin (Wuhan Huamei Bioengineering Co., Ltd.); N-hydroxysuccinimide (NHS, Shanghai Yanchang Biochemical Company); chloroauric acid (HAuCl 4 , Shanghai Reagent No. 1 Factory, China); trichloroethyl phosphate (TCEP), mercaptohexanol (MCH) (Aladdin Reagent Co., Ltd.).

[0047] The sequences of the peptides used (purity >95%, Bio-Thera Biotechnology) are as follows:

[0048] 11-mercaptodecanoic acid-GLSKKRPPPPPPGHKRT.

[0049] CHI660B electrochemical workstation (Shanghai Chenhua Instrument Office); MPI-E electrochemiluminescence analysis system (Xi'an Ruimai Analytical Instrument Co., Ltd.); three-e...

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Abstract

The invention belongs to the technical fields of analytical chemistry and electrochemical luminescence detection. The invention relates to an electrochemical luminescence sensor of cortex actin and a method for determining the cortex actin. A nanogold and graphene modified gold electrode is prepared by using electrochemical reduction, and peptide with an active group bipyridine ruthenium connection is self-assembled on the modified electrode, so that the electrochemical luminescence sensor is formed. When the cortex actin of a target object exists, the cortex actin is combined with the peptide to prevent thermolysin from cutting the peptide, and a strong electrochemical luminescence signal is generated; and otherwise, when no cortex actin exists, the thermolysin cuts the peptide, and the electrochemical luminescence signal is weakened. Therefore, the cortex actin is determined. The method has the characteristics of simplicity, high sensitivity, simple structure and low cost of an instrument device.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry and electrochemiluminescence detection, and particularly relates to a method for measuring cortical actin by using a peptide end-labeled with an electrochemiluminescence signal probe as a recognition element. technical background [0002] In the current biomedical research and clinical practice, it is an eternal requirement to establish new simple, fast and effective methods for protein detection. The most commonly used analysis and determination method at this stage is based on the combination of antibody recognition and signal amplifier. Although this method is effective, due to the complex preparation of antibodies and signal amplifiers, new methods need to be established to replace these traditional methods that require complex steps. [0003] In the present invention, peptides are used instead of antibodies as recognition units to simplify the assay steps. Peptide not only has ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N27/26G01N27/30
Inventor 混旭徐亚琼
Owner 石家庄卢米特生物科技有限公司
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