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Carassius auratus vitellogenin ELISA kit, preparation method and application thereof

An enzyme-linked immunosorbent adsorption and original protein technology, which is applied in the field of enzyme-linked immunosorbent assay kits, can solve problems such as endangering human health, wildlife hazards, and water environment pollution, and achieves the effect of a wide linear working range

Inactive Publication Date: 2009-04-01
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, a large number of pollutants enter the water environment, causing serious water pollution problems.
These chemicals have caused serious harm to wildlife; more importantly, these chemicals can enter the human food system along the food chain, seriously endangering human health

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0010] The preparation of the enzyme-linked immunosorbent assay (ELISA) kit for detecting goldfish vitellogenin is divided into the following steps:

[0011] (1) Preparation of pure goldfish vitellogenin

[0012] 10mg 17β-estradiol (17β-estradiol, E 2 ) was dissolved in 0.5mL of absolute ethanol, then 0.5mL of peanut oil was added, and mixed thoroughly to prepare 17β-estradiol emulsion. Inject goldfish with 17β-estradiol emulsion intraperitoneally to induce goldfish to secrete vitellogenin. The injection dose is 0.1mL / piece. After 10 days, blood is taken from the tail vein and centrifuged at 8000g at 4°C for 5 minutes to obtain vitellogenin-containing plasma. . Take 1 mL of the plasma and add it to a molecular sieve chromatography column, the filler is Sephacryl S-300, elute with 25mM Tris-HCl (pH7.5) containing 0.07M NaCl, collect the sample containing goldfish vitellogenin for ion exchange chromatography, The filler is DEAE Sepharose Fast Flow, and 25mM Tris-HCl containin...

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PUM

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Abstract

The invention discloses an enzyme-linked immunosorbent kit for detecting the vitellogenin of goldfish, and a preparation method and applications thereof. The preparation method comprises the following steps: when in preparation, firstly, the pure product of the vitellogenin of the goldfish is prepared; secondly, the polyclonal antibody of rabbit anti-vitellogenin of the goldfish is prepared; thirdly, the polyclonal antibody of the rat anti-vitellogenin of the goldfish is prepared; and fourthly, respectively one bottle of the pure product of the vitellogenin of the goldfish, the polyclonal antibody of the rabbit anti-vitellogenin of the goldfish and the polyclonal antibody of the rat anti-vitellogenin of the goldfish, one block of blank 96-hole enzyme-linked immunosorbent assay board and respectively one bottle of the serum of a negative contrasting group, coating liquid, confining liquid, sample diluent, washing liquid, color development liquid, terminator and goat anti-mouse secondary antibody marked by horse radish peroxidase are filled into a box body together, thus obtaining the enzyme-linked immunosorbent kit for detecting the vitellogenin of the goldfish. The kit of the invention can be applied to the survey that the internal secretion disturbs chemical substances and can quantitatively detect the vitellogenin in the blood of the goldfish, liver tissues and the culture solution of hepatic cells, with sensitivity, accuracy and convenience.

Description

technical field [0001] The invention relates to an enzyme-linked immunosorbent assay kit for detecting goldfish vitellogenin, a preparation method and application thereof. Background technique [0002] At present, a large number of pollutants enter the water environment, causing serious water pollution problems. These chemicals have caused serious harm to wild organisms; more importantly, these chemicals can enter the human food system along the food chain, seriously endangering human health. A growing body of research suggests that reduced fertility in wildlife, feminization of fish, increased cancer in humans, and reduced sperm quantity and quality are all strongly associated with exposure to endocrine-disrupting chemicals. This type of pollutants that can disrupt the endocrine system of humans and wild animals is collectively referred to as endocrine disrupting chemicals, which can interfere with the synthesis, release, transportation, metabolism, and binding and functio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
Inventor 邴欣汝少国潘宗保
Owner OCEAN UNIV OF CHINA
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