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Competitive inhibiting enzyme-linked immune chip kit for detecting obesity factors and preparation method of competitive inhibiting enzyme-linked immune chip kit

A competitive inhibition, enzyme-linked immunosorbent technology, applied in the field of biomedicine, can solve the problems of low sensitivity, cumbersome operation, single detection index, etc., and achieve the effect of high sensitivity, specificity and good repeatability.

Active Publication Date: 2011-11-09
RAYBIOTECH INC GUANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at 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 obesity factor detection kit, which can simultaneously The detection of eleven obesity factors overcomes the shortcomings of the existing technology such as cumbersome operation, single detection index, expensive equipment, and low sensitivity. It is cheap, convenient, sensitive, accurate, high-throughput, and requires less specimens. Advantages of laboratory promotion and scale

Method used

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  • Competitive inhibiting enzyme-linked immune chip kit for detecting obesity factors and preparation method of competitive inhibiting enzyme-linked immune chip kit
  • Competitive inhibiting enzyme-linked immune chip kit for detecting obesity factors and preparation method of competitive inhibiting enzyme-linked immune chip kit
  • Competitive inhibiting enzyme-linked immune chip kit for detecting obesity factors and preparation method of competitive inhibiting enzyme-linked immune chip kit

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Example 1: Preparation of a competitive inhibitory ELISA kit for detecting obesity factors.

[0034] In order to detect whether there are corresponding obesity factors in the sample, the bottom membrane immobilized with antibodies specific to the following obesity factors: visfatin, neuropeptide Y, angiotensin II, resistin, adiponectin, retinoid Alcohol-binding protein 4, vascular adhesion protein, transcriptional regulatory peptide, antigen-presenting cells, growth hormone receptor, and neuron-specific enolase.

[0035] 1. Antibody preparation:

[0036] Using specific antibodies against the obesity factors listed in Table 1, the source, concentration, and protein names of the antibodies are detailed in Table 1:

[0037]Table 1 The name of the obesity factor targeted by the specific antibody, the source and concentration information of the antibody.

[0038]

[0039] [0026] 2. Spotting and sealing

[0040] Spot 50-500nl of tris buffer containing 0.5-100ng of sp...

Embodiment 2

[0042] Embodiment 2: The experiment of detecting obesity factors with the kit of the present invention.

[0043] 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 dot matrix is ​​divided into mutually independent reaction areas. In this embodiment, 8 square boxes are provided, which are produced by Ruiboao Biotechnology Co., Ltd. of the United States. 2 ml of blocking solution is added to each square and placed Incubate at room temperature for 30 minutes, and then perform the following steps in sequence:

[0044] 1. Add sample

[0045] Experiment 1: Using the kit of the present invention to detect obesity factors in human serum samples.

[0046] Aspirate the blocking solution in each square, put 100 μl ~ 5ml of the sample diluted with the blocking solution into the square with a membrane, and then sha...

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Abstract

The invention discloses a competitive inhibiting enzyme-linked immune chip kit for detecting obesity factors. The kit comprises (1) a basement membrane, and (2) a reactant and a detection agent, wherein a plurality of specific antibodies are simultaneously fixed on the basement membrane, the specific antibodies and the obesity factors can undergo antibody-antigen reaction, and the specific antibodies are respectively fixed on the basement membrane to form a plurality of independent identification sites; and the reactant and the detection agent are used for detecting whether a substance capable of undergoing the antibody-antigen reaction with the specific antibodies exists in a sample to be detected by a competitive inhibiting enzyme-linked immune chip method. The invention also discloses a preparation method of the kit. The method comprises a step of fixing the specific antibodies on the basement membrane. The kit can be used for simultaneously detecting multiple obesity factors by adopting a competitive inhibiting enzyme-linked immune chip technology, overcomes multiple defects of the prior art, and has the advantages of low price, convenience, sensitivity, accuracy, high flux, low specimen consumption, capability of being popularized in a common laboratory and large scale and the like.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a competitive inhibitory ELISA kit for detecting obesity factors. technical background [0002] The traditional view holds that the role of adipose tissue is single and passive. However, with the discovery of the first protein complement D secreted by adipocytes in 1987; especially in 1994, the gene for adipocyte-specific secretion of protein leptin was successfully cloned, and after leptin was found to be closely related to obesity, its important secretion function attracted more and more attention. So far, more than 20 biologically active substances such as complement D, leptin, and resistin have been isolated from adipose tissue, including cytokines, growth factors, enzymes, hormones, complement factors, and matrix proteins, collectively referred to as obesity factors. or adipocytokines. Experiments have shown that obesity factor acts on multiple organs through differen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/544G01N33/545G01N33/531
Inventor 黄若磐
Owner RAYBIOTECH INC GUANGZHOU
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