Human adiponectin enzyme-linked immunosorbent detection kit, preparation method and application thereof

A technology of enzyme-linked immunosorbent adsorption and detection kits, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of inconvenient use and operation, long detection time, high price of ELISA kits, etc., and achieve low cost, The effect of being convenient for clinical mass use and convenient operation

Inactive Publication Date: 2009-11-04
袁洪 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing ELISA kits are expensive, the operation is not convenient, and the detection takes a long time

Method used

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  • Human adiponectin enzyme-linked immunosorbent detection kit, preparation method and application thereof
  • Human adiponectin enzyme-linked immunosorbent detection kit, preparation method and application thereof
  • Human adiponectin enzyme-linked immunosorbent detection kit, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The APN quantitative ELISA kit of the present embodiment comprises: a microtiter plate coated with an anti-human APN monoclonal antibody (MAB10651), standard dry powder, detection antibody, enzyme-labeled avidin, sample reagent diluent, TMB enzyme Substrate chromogenic solution, concentrated washing solution and termination reaction solution. in:

[0035] The standard freeze-dried powder is the recombinant gAPN protein standard;

[0036] The detection antibody is biotin-labeled anti-human APN monoclonal antibody (MAB1065);

[0037] Enzyme-labeled avidin is HRP-streptomycin, which is packaged for Invitrogen products;

[0038] TMB substrate chromogenic solution is sub-packaged for KPL products;

[0039] The sample diluent is 10mM pH7.4 phosphate buffer containing 0.05% Tween-20;

[0040] The concentrated washing solution is 0.1M pH7.4 phosphate buffer containing 0.05% Tween-20;

[0041] The stop reaction solution was 2 M H 2 SO 4 ;

[0042] The stated percentages ...

Embodiment 2

[0087] Embodiment 2: the analytical precision (CV%) of kit of the present invention

[0088] Three clinical specimen serums were randomly selected, and the APN concentration in the sample and the analytical precision of the method were measured and calculated by using the above-mentioned APN ELISA kit. The test results are shown in Table 1:

[0089] Table 1 Kit precision test of the present invention

[0090] serum sample

[0091] The detection results show that the detection precision of the kit of the present invention meets the standard requirements of the ELISA quality index, and the analytical sensitivity is 150ng / ml, which is suitable for actual clinical detection.

Embodiment 3

[0092] Embodiment 3: the methodological comparison of kit of the present invention

[0093] Get 30 clinical serum specimens at random, apply the kit of the present invention and the people's APN concentration ELISA quantitative detection kit of Shenzhen Enuojin Company to detect simultaneously, compare the correlation of the two; the result both correlation is good, correlation coefficient r= 0.935, P2 =0.8737 (see Figure 5 ).

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Abstract

The invention provides a human adiponectin enzyme-linked immunosorbent detection kit, a preparation method and application thereof. The kit comprises antihuman APN monoclonal antibody-coated ELISA plates, recombinant gAPN protein standard, antihuman APN monoclonal detection antibodies, HRP-streptomycin, 10 mM of phosphate buffer with a pH of 7.4, TMB enzyme-substrate chromogenic solution, 0.1 M of phosphate buffer with a pH of 7.4 and 2 M of H2SO4. The invention also comprises the preparation method of the kit and the application of the kit in the detection of human-blood APN content. The kit has the advantages that the kit is convenient and fast to use and operate, capable of getting detection results within 2 hours, high in specificity, low in cost and convenient for clinical large-scale use.

Description

technical field [0001] The invention relates to a human adiponectin enzyme-linked immunosorbent assay kit and its preparation method and application, in particular to an enzyme-linked immunosorbent assay (ELISA) kit for detecting adiponectin in human serum or plasma and its preparation Methods and applications. Background technique [0002] Adiponectin (APN) is an important plasma hormone protein first discovered by Scherer et al. in the mid-1990s (Scherer PE, WilliamsS, FoglianoM, 1995, A novel serum protein similar Clqproduced exclusively in adipocytes.J Biol Chem, 270:26746-26749). APN, also known as Acrp30, AdipoQ or GBP28, is a specific plasma protein secreted by adipose tissue. It is abundant in the circulation, with a serum concentration of 5-30 mg / L, accounting for about 0.01% of the total human serum protein. Serum protein levels such as cortisol and interleukin-6 were 103-106 times higher (Shimada K, Miyazaki T, Daida H, 2004, Adiponectin and atherosclerotic dise...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543G01N33/535
Inventor 袁洪李登清陈琳磊何笑恬曹琳金维荣
Owner 袁洪
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