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Human interleukin 35 enzyme-linked immunosorbent assay kit and preparation method thereof

An enzyme-linked immunosorbent and interleukin technology, applied in the biological field, can solve the problem of less detection tools, and achieve the effects of no special protection, simple experimental operation and high stability.

Active Publication Date: 2016-05-04
WUHAN CLOUD CLONE CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Because IL-35 is a newly discovered cytokine, there are few detection tools for it on the market

Method used

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  • Human interleukin 35 enzyme-linked immunosorbent assay kit and preparation method thereof

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Embodiment Construction

[0039] Describe technical content of the present invention in detail below by embodiment:

[0040] The main technical route is as follows:

[0041] Firstly, according to the sequences of IL-12p35 and IL-27EBI3 in the NCBI gene bank Genebank, primers for amplifying the corresponding fragments and primers required for linking IL-12p35 and IL-27EBI3 were designed respectively; The corresponding gene fragments were called out from the library. Among them, IL-12p35 and IL-27EBI3 are used to immunize animals, and specific antibodies against the two fragments are prepared respectively;

[0042] Full-length IL-35 protein was used as a standard in the IL-35 ELISA kit.

[0043] 1. Preparation of recombinant IL-12p35 protein and monoclonal antibody:

[0044] 1) IL-12p35 primer design and gene fragment amplification

[0045]For the amplification of the IL-12p35 gene fragment, the reference gene sequence is Genebank No. NM_000882.3. According to the found genes, the upstream and downs...

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Abstract

The invention discloses a human interleukin-35 enzyme-linked immunosorbent assay kit and a preparation method thereof. The kit includes a mouse anti-IL-12p35 monoclonal antibody (coating antibody), a full-length recombinant protein IL- 35 (standard), biotinylated mouse anti-IL-27EBI3 monoclonal antibody (detection solution A), substrate TMB and stop solution. The kit of the invention has the advantages of high stability and low toxicity, and the experimental operation is simpler than the RIA method and the mass spectrometry method, does not require special protection and has a short period, and can meet the requirements of ordinary laboratories.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a human interleukin-35 enzyme-linked immunosorbent assay kit and a preparation method thereof. Background technique [0002] Interleukin 35 (IL35), a newly discovered member of the IL-12 cytokine family, is a heterodimer composed of two subunits, namely EBI3 and IL-12p35. EBI3 is induced by Epstein-Barr virus-infected B lymphocytes and is encoded as a 34kDa glycoprotein, 27% of which is identical to IL-12p40 in amino acids; IL-12p35 is homologous to other single-chain cytokines such as IL-6. IL-35 was discovered in 1997 until 2007 when Collison et al first described the composition and function of IL-35, and found that IL-35 is secreted by regulatory T-cells (Treg) and regulates immune function important cytokines. Treg cells are an important subset of CD4+ T cells, which play a fundamental role in maintaining self-tolerance, preventing autoimmune diseases, limiting chr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/544
Inventor 李华渊孙颖姚雯何峰容杨娟汪景万定一李庆
Owner WUHAN CLOUD CLONE CORP