Unlock instant, AI-driven research and patent intelligence for your innovation.

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

An enzyme-linked immunosorbent assay and interleukin technology, applied in the biological field, can solve the problem of fewer detection tools, and achieve the effect of no special protection, high stability, and short cycle

Active Publication Date: 2013-10-16
WUHAN CLOUD CLONE CORP
View PDF10 Cites 0 Cited by
  • 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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Human interleukin 35 enzyme-linked immunosorbent assay kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

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 downst...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a human interleukin 35 enzyme-linked immunosorbent assay kit and a preparation method thereof. The kit comprises a mouse anti-IL-12p35 monoclonal antibody (namely a coated antibody), a full length recombinant protein IL-35 (namely a standard substance), a biotinylated mouse anti-IL-27EBI3 monoclonal antibody (namely a detection solution A), a substrate TMB and a stop solution. The kit has the advantages of high stability and low toxicity; moreover, the experimental operation is simpler compared with that in radioimmunoassay (RIA) and mass-spectrography, special protection is not required, the period is short, and the requirements of general laboratories can be met.

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 plays a role in immune function. An important cytokine that regulates effects. Treg cells are an important subset of CD4+ T cells, which play a fundamental role in maintaining self-tolerance, preventing a...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/544
Inventor 何峰容杨娟汪景彭夫望孙颖高强李庆姚雯
Owner WUHAN CLOUD CLONE CORP