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Immunofluorescence kit and application for detection of eggshell precursor protein of flounder flounder

A precursor protein and immunofluorescence technology, applied in the field of environmental detection, can solve the problems of lack of location and quantitative detection of eggshell precursor protein, obtain eggshell precursor protein, and not establish eggshell precursor protein, etc., to improve detection sensitivity The effect of high specificity and low background value

Inactive Publication Date: 2021-07-30
OCEAN UNIV OF CHINA
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Problems solved by technology

Therefore, eggshell precursor protein is a more sensitive biomarker for environmental estrogen screening. However, no detection kit for eggshell precursor protein has been established at home and abroad. The main reason is that traditional separation and purification methods are difficult to obtain from fish. Eggshell precursor protein cannot provide high-purity antigens for the development of immunoassay technology, so no detection method for eggshell precursor protein has been established yet, and there is a lack of methods for the localization and quantitative detection of eggshell precursor protein production sites

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  • Immunofluorescence kit and application for detection of eggshell precursor protein of flounder flounder
  • Immunofluorescence kit and application for detection of eggshell precursor protein of flounder flounder
  • Immunofluorescence kit and application for detection of eggshell precursor protein of flounder flounder

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Embodiment Construction

[0023] An immunofluorescent kit for detecting eggshell precursor protein of brown flounder, characterized in that the kit is equipped with one FITC-labeled mouse polyclonal antibody against eggshell precursor protein of brown flounder.

[0024] The immunofluorescence detection technique for preparing and detecting the eggshell precursor protein of brown teeth comprises the following steps:

[0025] (1) Preparation of pure flounder eggshell precursor protein;

[0026] (2) Preparation of polyclonal antibody against eggshell precursor protein of flounder flounder;

[0027] (3) Preparation of FITC-labeled polyclonal antibody to eggshell precursor protein of brown flounder;

[0028] Wherein, the (1) method for preparing pure flounder eggshell precursor protein is as follows:

[0029] (1) Preparation of pure flounder eggshell precursor protein

[0030] ① Construction of prokaryotic expression vector for eggshell precursor protein

[0031]A large amount of Chg H mRNA was induced ...

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Abstract

Immunofluorescence kit and application for detection of eggshell precursor protein of flounder flounder. The eggshell precursor protein of brown flounder flounder was prepared by prokaryotic expression, the polyclonal antibody of eggshell precursor protein of brown flounder was developed by immunizing mice, and FITC was labeled on the purified polyclonal antibody; Construction of polyclonal antibody against eggshell precursor protein of flounder flounder and immunofluorescence kit for detection of eggshell precursor protein of brown flounder. The invention not only can be used to detect the production of the egg shell precursor protein in the brown flounder fish body, but also can locate the position of the egg shell precursor protein in the liver tissue. Compared with the previous immunohistochemistry, the present invention has fewer steps, saves time, detects the background value, and provides an important tool for the screening and detection of estrogen in marine environment and ecological environment risk assessment.

Description

technical field [0001] The invention belongs to the field of environmental detection, and in particular relates to an immunofluorescence kit for detecting eggshell precursor protein of flounder flounder and its application. Background technique [0002] Environmental estrogen, as a type of exogenous compound, can interfere with the synthesis, release, movement, metabolism, and combination of normal secreted substances in the body after entering the body, activate or inhibit the function of the endocrine system, thereby destroying its role in maintaining the stability and regulation of the body . In recent years, estrogen pollution in the environment poses a potential threat to the ecological balance of the natural environment and human health. After environmental estrogen substances enter the human body, women often suffer from uterine fibroids, ovarian cancer and other diseases, and men often suffer from testicular cancer, Decreased sperm count and quality. Estrogenol bel...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/74G01N33/68G01N33/533
Inventor 王军张振忠汝少国王蔚
Owner OCEAN UNIV OF CHINA
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