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Protein chip kit for detecting inflammatory factors and preparation method thereof

A protein chip and inflammatory factor technology, applied in the field of biomedicine, can solve the problems of low sensitivity, cumbersome operation, expensive instruments, etc., achieve the effect of less sample consumption and overcome cumbersome operation

Active Publication Date: 2013-08-28
RAYBIOTECH INC GUANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In view of the deficiencies in the prior art, the technical problem to be solved by the present invention is to provide a high-throughput, high-sensitivity, high-specificity and low-cost inflammatory factor detection kit, which can detect forty inflammatory factors by using protein chip technology , which overcomes the defects of the existing technology such as cumbersome operation, single detection index, need for expensive instruments, and low sensitivity, and has the advantages of cheap, convenient, sensitive, accurate, high-throughput, less specimen consumption, and can be popularized and scaled in ordinary laboratories Etc

Method used

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  • Protein chip kit for detecting inflammatory factors and preparation method thereof
  • Protein chip kit for detecting inflammatory factors and preparation method thereof
  • Protein chip kit for detecting inflammatory factors and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Preparation of a protein chip kit for detecting inflammatory factors.

[0028]In order to detect whether there are corresponding inflammatory factors in the sample, the bottom membrane immobilized with specific antibodies against the following proteins: eotaxin (EOTAXIN), leukocyte inducer-2 (EOTAXIN-2), granulocyte Colony-stimulating factor, (GCSF), recombinant human granulomacrophage colony-stimulating factor (GM-CSF), intercellular adhesion molecule-1 (ICAM-1), interferon-γ (IFN-γ), activated human T cells A gene product (I-309), interleukin-1α (IL-1α), interleukin-1β (IL-1β), interleukin-2 (IL-2), interleukin-3 (IL -3), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-6sR (IL-6sR), interleukin-7 (IL-7), interleukin IL-8 (IL-8), interleukin-10 (IL-10), interleukin-11 (IL-11), interleukin-12p40 (IL-12p40), interleukin-12p70 (IL- 12p70), interleukin-13 (IL-13), interleukin-15 (IL-15), interleukin-16 (IL-16), interleukin-17 (IL-17), chemokines Inter...

Embodiment 2

[0040] Example 2: The experiment of detecting inflammatory factors with the kit of the present invention.

[0041] The bottom film is placed in the matching square box. Since there are a plurality of chip lattices distributed on the bottom film of this embodiment, the square box is provided with 8 square grids in this embodiment. Each chip array is divided into mutually independent reaction areas. In this example, 8 square boxes are provided and made by RayBiotech. After adding 2 ml of blocking solution to each square, place it at room temperature and incubate for 30 minutes. Then perform the following steps in sequence:

[0042] 1. Add sample

[0043] Aspirate the blocking solution in each square, put 100 microliters to 5 ml of the sample diluted with the blocking solution into the square with a membrane, and then shake it on a shaker at room temperature for 1 to 2 hours, or in React at 4°C for 12-18 hours.

[0044] 2. Membrane washing

[0045] Washing solution I cleaning...

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Abstract

The invention discloses a protein chip kit for detecting inflammatory factors, which comprises a basement membrane (1) and a reactant and a detection agent (2). A plurality of kinds of specific antibodies are fixed on the basement membrane; the specific antibodies and the inflammatory factors can undergo an antigen-antibody reaction and each specific antibody is respectively fixed on the basementmembrane for forming a plurality of independent recognition loci; and the reactant and the detection agent are used for detecting whether substances capable of undergoing the antigen-antibody reaction with the specific antibodies exist in a sample to be detected or not by an array-ELISA method. The invention also discloses a method for preparing the protein chip kit for detecting the inflammatoryfactors, which comprises a step of fixing the specific antibodies on the basement membrane. The kit of the invention adopts protein chip technology, can detect 40 inflammatory factors, overcomes a plurality of shortcomings and has the advantages of low cost, convenient use, high sensitivity and accuracy, high flux, small using amount of samples, capability of being popularized in an ordinary laboratory, massive production and the like.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a protein chip kit for detecting inflammatory factors. technical background [0002] Inflammation is an autogenous protective mechanism for organisms to eliminate harmful stimuli and promote healing. The process involves complex biochemical reactions. These harmful stimuli include pathogens, damaged cells or irritants, etc. Inflammation generally manifests as acute inflammation and chronic inflammation, among which, acute inflammation is the body's initial response to harmful stimuli, which is achieved by increasing the plasma volume and white blood cell count of damaged tissues, and is accompanied by a series of chain biological reactions, including local microcirculation , the immune system and various cells in damaged tissues; while chronic inflammation will lead to gradual changes in the cells of the inflammation site, which is a process of coexistence of injury and heali...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/544
Inventor 黄若磐
Owner RAYBIOTECH INC GUANGZHOU
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