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Tri-antibody sandwich ELISA detection method of IgG of tree shrew

A detection method and three-antibody technology, applied in the field of immunological detection, can solve the problems of animal model obstacles and difficult realization.

Inactive Publication Date: 2013-12-18
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Antibody research is difficult to achieve. The application of tree shrews to human diseases, especially animal models of viruses, is a big obstacle. Therefore, the preparation of tree shrew antibody research tools and the establishment of antibody methods have become urgent problems to be solved.
So far, there is no report on the method for detecting tree shrew IgG in the prior art

Method used

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  • Tri-antibody sandwich ELISA detection method of IgG of tree shrew
  • Tri-antibody sandwich ELISA detection method of IgG of tree shrew

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Embodiment 1

[0029] The ELISA plates used in the following three-antibody sandwich ELISA method of the present invention are all 96-well single-well detachable ELISA plates purchased from Coster Company, and the coating buffer is 0.05mol / L carbonate buffer at pH9.6 solution, the sample diluent was PBS buffer at pH 7.4, the washing solution was 0.05% PBST buffer, and the commercial goat anti-rabbit IgG-HRP was purchased from Boster Bioengineering Co., Ltd.

[0030] Establishment of triple antibody sandwich ELISA method:

[0031] 1. Method:

[0032] 1.1. Establishment of IgG standard curve:

[0033]Coating solution containing 1 / 400 mouse anti-tree shrew IgG monoclonal antibody ascites to coat the microtiter plate, 100 μl / well, incubate at 37°C for 2 hours, and overnight at 4°C; wash 3 times with PBST, 300 μl / well, micro-shock for 3 minutes each time , discard the washing solution and pat dry on absorbent paper; block with 2% BSA, 200μl / well, incubate at 37°C for 2h; wash with PBST 3 times,...

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Abstract

The invention discloses a tri-antibody sandwich ELISA (enzyme-linked immunosorbent assay) detection method of the IgG (Immunoglobulin G) of a tree shrew, which is applicable to the detection of the total content of IgG in the blood plasma of the tree shrew. The method comprises the following steps: coating an enzyme-label plate with the monoclonal antibody in the anti-rat IgG of the tree shrew as a capture antibody, and combining with the IgG of the tree shrew to be detected; and establishing the tri-antibody sandwich detection method of the IgG of the tree shrew by optimizing the concentration of the antibody and other multi-factor conditions by using the polyclonal antibody of the anti-rabbit IgG of the tree shrew as the secondary antibody and the commercial goat anti-rabbit IgG-HRP as the detection antibody. In the invention, the IgG level of the blood plasma of 30 tree shrews is detected by using the method, wherein according to the detection, the normal IgG level of the blood plasma of the tree shrews is 8.6g / L-17.8g / L. The tri-antibody sandwich ELISA detection method of the IgG of the tree shrew provides necessary tools for the research on the tree shrew as the animal model for the experiment on human diseases and the immunity mechanism of the relates diseases.

Description

Technical field: [0001] The invention belongs to the field of immunological detection methods, in particular to an antibody sandwich ELISA method for detecting tree shrew IgG. Background technique: [0002] Tree shrew (Tupaia, tree shrew) is a small squirrel-like climbing mammal, mainly distributed in tropical and subtropical regions of southeastern Asia, and widely distributed in Yunnan, Guangxi, Guangdong and Hainan in my country. Tree shrews are small in size, fast in growth and reproduction, easy to catch, domesticated, and low in breeding costs. Their metabolic characteristics and anatomical features are closer to humans than rodents. Many people classify tree shrews as primates. With the gradual reduction of non-human primate resources at home and abroad and the development trend of miniaturization of experimental animals, tree shrews have attracted widespread attention as a possible animal model for the study of human-related diseases, and have been used in virology, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/543G01N33/558
Inventor 夏雪山温志刚冯悦刘丽魏大巧杨恒
Owner KUNMING UNIV OF SCI & TECH
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