Leptin detection kit

A detection kit and leptin technology, applied in the fields of biology and medical testing, can solve the problems of poor sensitivity and repeatability, and achieve the effect of high sensitivity and good repeatability

Inactive Publication Date: 2013-01-30
ANHUI ANKE BIOTECHNOLOGY (GRP) CO LTD
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0003] Leptin detection kit is a common means of serological diagnosis of leptin, which is carried out by applying the basic principle of enzyme-linked immunosorbent assay, which is also called enzyme-linked immunosorbent assay (enzyme-linked immunosorbent assay), Abbreviated as ELLSA, the ELLSA process includes the adsorption of antigen (antibody) on a solid phase carrier called coating, adding the antibody (antigen) to be tested, and then adding the corresponding enzyme-labeled antibody (antigen) to generate antigen (antibody)-antibody to be tested (antigen) )-enzyme-labeled antibody complex, and then react with the substrate of the enzyme to generate a colored product. The amount of the antibody (antigen) is calculated by means of the light absorption of the spectrophotometer. The quantification of the antibody (antigen) to be tested is proportional to the color production. The leptin detection kit is one of the auxiliary diagnostic means for clinical obesity and diabetes in the prior art, but the sensitivity and repeatability of the existing leptin enzyme-labeled reagents are poor

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Embodiment Construction

[0024] step 1

[0025] The composition of the leptin detection kit:

[0026] (1) 5 bottles of leptin reference products with different concentrations and specifications: 0ng / ml, 1ng / ml, 10ng / ml, 25ng / ml, 50ng / ml, 1-3ml / bottle;

[0027] (2) Coating antibody microplates;

[0028] (3) Enzyme-labeled anti-leptin;

[0029] (4) Washing solution, 20ml / bottle;

[0030] (5) Both chromogenic solution A and chromogenic solution B are 10-15ml / bottle;

[0031] (6) Stop solution, 15-20ml / bottle.

[0032] step 2

[0033] The preparation of recombinant human leptin polypeptide antigen monoclonal antibody comprises the following steps:

[0034] (1) Immunize female Balb / c mice for 6-8 weeks with recombinant human leptin polypeptide antigen;

[0035] (2) Take the immune spleen cells of the above mice and myeloma cells SP2 / 0, and fuse them under the action of 50% polyethylene glycol to obtain hybridoma cells; polyethylene glycol is produced by Sigma in the United States, with a molecular w...

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Abstract

The invention relates to a leptin detection kit. The leptin detection kit comprises a leptin reference product, an antibody-coated microtiter plate, an enzyme-labeled anti-leptin, a washing solution, a color development solution and a stop solution, wherein the leptin reference product is of a recombinant human leptin polypeptide antigen, and the antibody-coated microtiter plate and the enzyme-labeled anti-leptin are of recombinant human leptin polypeptide monoclonal antibodies. The detection principle of the leptin detection kit disclosed by the invention is as follows: the recombinant human leptin polypeptide monoclonal antibody is taken as the antibody-coated microtiter plate, a serum sample to be detected is added, the corresponding enzyme-labeled antibody is further added, a compound of the coating antibody, the serum sample to be detected and the enzyme-labeled antibody is generated, the color development solution is added for color development, then the stop solution is further added for stopping, the residual solution is washed away by the washing solution, the light absorption value of the serum sample to be detected is determined, and the leptin content in the serum sample to be detected can be obtained by comparing with a standard curve.

Description

technical field [0001] The invention relates to the fields of biology and medical examination, in particular to a leptin detection kit. Background technique [0002] With the improvement of human living standards, obesity has become a common problem in the world today. Obesity is a disorder of food intake and energy consumption balance mechanism, which can cause various diseases, such as type II diabetes, hypertension, hyperlipidemia and cancer. Leptin is often referred to as obesity protein, which is the product of obesity genes. Leptin acts on the hypothalamus, controls metabolism, energy consumption and reproductive system, and regulates body fat content and glucose metabolism. Exclusively expressed in adipocytes, circulating leptin levels are positively correlated with body fat mass. Leptin plays a role in hematopoiesis and adrenal cortex function. Normal human serum leptin levels are low, and changes in leptin may occur in arteriosclerosis, unstable growth hormone, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
Inventor 宋礼华徐振山杜贤宇周业亭宋社吾饶海军
Owner ANHUI ANKE BIOTECHNOLOGY (GRP) CO LTD
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