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Enzyme-linked immune kit for detecting copper ion and application thereof

An enzyme-linked immunosorbent reagent and copper ion technology, which can be used in measurement devices, instruments, scientific instruments, etc., can solve the problems of difficulty in adapting to the rapid customs clearance of product import and export, and inability to use on-site testing, so as to solve food safety problems and have no radioisotopes. The effect of pollution and error reduction

Inactive Publication Date: 2012-09-19
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional heavy metal copper detection methods such as: atomic absorption spectrometry, inductively coupled plasma emission spectrometry, etc. require large and expensive analytical instruments, which cannot be used for on-site detection, and are difficult to adapt to the environment and on-site random inspection of agricultural and livestock products and rapid customs clearance of product import and export. requirements

Method used

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  • Enzyme-linked immune kit for detecting copper ion and application thereof
  • Enzyme-linked immune kit for detecting copper ion and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The ELISA kit for detecting copper ions in this embodiment includes copper ion-specific antibodies, copper ion-carrier protein conjugates and enzyme-labeled secondary antibodies, and is also provided with coated copper ion antigens or anti-copper ion-specific antibodies ELISA plate, copper ion standard solution, substrate chromogenic reagent, washing solution, stop solution and concentrated sample diluent.

[0037] The preparation method of each raw material of the kit of the present embodiment is as follows:

[0038] 1. Preparation and identification of complete antigen:

[0039] Dissolve 0.5-10 mg of water-soluble copper salt in 50 μL of ultrapure water to prepare 2.0-80 mg / mL of Cu 2+ solution; take 0.8-20mg bifunctional chelating agent p-SCN-Bn-DOTA and dissolve it in 50μL dimethyl sulfoxide to prepare a 16-400mg / mL p-SCN-Bn-DOTA solution; take 2.0-20.0mg carrier protein molybdenum Pore ​​hemocyanin (KLH) and bovine serum albumin (BSA) were dissolved in 1mL HEPES ...

Embodiment 2

[0054] The preparation method of the enzyme-linked immunosorbent assay kit that detects copper ion, it comprises following components:

[0055] (1) A microtiter plate coated with a copper ion-coupled antigen;

[0056] (2) Goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0057] (3) Copper ion single antibody concentrate;

[0058] (4) 6 bottles of copper ion standard solution, the concentrations are 0.25mmol / L, 0.125mmol / L, 0.0625mmol / L, 0.03125mmol / L, 0.015625mmol / L, 0.0078125mmol / L.

[0059] (5) The substrate chromogenic solution is composed of A liquid and B liquid, the substrate chromogenic liquid A liquid is carbamide peroxide, and the substrate chromogenic liquid B liquid is tetramethylbenzidine;

[0060] (6) The stop solution is 2mol / L hydrochloric acid or sulfuric acid;

[0061] (7) The washing liquid is 0.01M, the pH is 7.4, and contains 0.05%-0.5% Tween-20 phosphate buffer;

[0062] (8) The concentrated sample solution is 0.1% Tween-20 phosphate...

Embodiment 3

[0064] Detection of residual copper ions in samples

[0065] Sample pretreatment

[0066] Accurately measure 5ml of milk into a 100ml beaker. Add 20ml HCl, let stand for 1h, centrifuge at 4000rpm for 15min, take the supernatant, add chelating agent, let stand for 0.5h, centrifuge at 4000rpm for 15min, take the supernatant.

[0067] Detection with a kit

[0068] Add 50 μl of a series of standard or sample solutions to the microwells of the copper ion-chelating agent-BSA conjugate-coated microtiter plate, then add 50 μl of antibody working solution, and react at room temperature for 1 h. Pour out the liquid in the wells, add 300 μl washing solution to each well, pour out the liquid in the wells after 30 seconds, repeat this operation for a total of 3 washes, and pat dry with absorbent paper. Add 100 μl of enzyme-labeled anti-antibody to each well, and incubate at room temperature in the dark for 30 min. Substrate chromogenic solution A (carbamide peroxide) 50 μl, add B solut...

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Abstract

The invention relates to an enzyme-linked immune kit for detecting copper ions and application thereof. The kit comprises a copper ion specific antibody, a copper ion-carrier protein coupler and an enzyme-labeled second antibody, and is further provided with an enzyme labeled plate coated by copper ion antigen or the anti-copper ion specific antibody, a copper ion standard solution, a substrate developer, washing liquid, terminating liquid and concentrated sample diluent. The enzyme-linked immune detection kit for the copper ions provided by the invention has the advantages of sensitivity, quickness and accuracy and is mainly used for screening of a large amount of samples; main reagents in the kit are provided in the form of operating liquid, have the characteristics of convenience for use, high specificity, high flexibility, high precision, high accuracy and on the like, and can quickly detect the residual copper ions in water and animal products.

Description

technical field [0001] The invention relates to an enzyme-linked immunosorbent reagent kit for detecting copper ions, and also relates to an application of the test kit in detecting copper ion content in a sample, belonging to the field of enzyme-linked immunosorbent detection. Background technique [0002] Copper accumulates in the soil and crops, which will pollute grain grains, and copper will remain in agricultural and livestock products to varying degrees, resulting in excessive copper pollution in agricultural and livestock products, seriously affecting the quality and international competitiveness of my country's agricultural and livestock products, and It also seriously endangers human health. Therefore, all countries in the world attach great importance to the control and monitoring of heavy metal pollution in the environment and agricultural products. Traditional heavy metal copper detection methods such as: atomic absorption spectrometry, inductively coupled plasm...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531G01N33/535
Inventor 孔涛杨自军赵振升张才郝雪琴周变华常景周李润乐魏迎军李嘉嘉王振华
Owner HENAN UNIV OF SCI & TECH
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