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Human serum CCL2 enzyme-linked immunosorbent assay kit as well as preparation and use methods thereof

An enzyme-linked immunosorbent assay and human serum technology, applied in the field of immunological detection, can solve the problems of insufficient sensitivity and accuracy for clinical application, unstable kit quality, unsuitable time-consuming operation, etc., and improve the level of clinical diagnosis and treatment. , low cost, short time effect

Inactive Publication Date: 2014-12-10
广西南宁隆吉维特生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are ELISA kits for the determination of CCL2 on the market abroad, but most of them are only for scientific research applications. Due to factors such as high price and time-consuming operation, they are not suitable for clinical use, such as the Immunoassay DCP00 series of R&D Systems
However, domestic ELISA kits for the determination of CCL2 are moderately priced, but the quality of the kits is not stable, and the sensitivity and accuracy cannot meet the requirements of clinical applications.

Method used

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  • Human serum CCL2 enzyme-linked immunosorbent assay kit as well as preparation and use methods thereof
  • Human serum CCL2 enzyme-linked immunosorbent assay kit as well as preparation and use methods thereof
  • Human serum CCL2 enzyme-linked immunosorbent assay kit as well as preparation and use methods thereof

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Experimental program
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preparation example Construction

[0037] Preparation of the above-mentioned CCL2 standard: take recombinant human CCL2 / MCP-1 (279-MC-010) as a standard, take 25 μg of recombinant human CCL2 / MCP-1 (279-MC-010) and add 1 ml of 1% BSA Fully dissolve the PBS solution to obtain a standard solution with a liquid concentration of 10 μg / ml, dilute it 5000 times to 2000 pg / ml, take 2000 pg / ml as the highest point concentration, and then do 2-fold dilution, and dilute 6 gradients in total to obtain 1000pg / ml, 500pg / ml, 250pg / ml, 125pg / ml, 62.5pg / ml and 31.25pg / ml, in addition, take the dilution of the standard as the lowest concentration, ie 0pg / ml. The diluent of the standard here is phosphate buffer saline (PBS) containing 0.1% bovine serum albumin (BSA) at pH 7.2-7.4.

[0038] Preparation of the above-mentioned biotin-labeled anti-human CCL2 antibody: using biotinylated human CCL2 / MCP-1 antibody (BAF279) as the second antibody, the stock solution concentration is 50 μg / ml, and pH7.2-7.4 containing 0.1% bovine serum w...

Embodiment 1

[0063] (1) Some important conditions of the human serum CCL2 ELISA kit are as follows: coat with mouse anti-human CCL2 monoclonal antibody at a concentration of 2000ng / ml, and the concentration of biotin-labeled anti-human CCL2 antibody is 125ng / ml;

[0064] (2) Test sample introduction: Group 1 is 8 breast cancer samples for comparison, Group 2 is 16 samples positive for tumor metastasis, Group 3 is 16 samples negative for tumor metastasis, and Group 4 is physical examination of Huanghe Hospital Control 8 copies;

[0065] (3) Adding samples: Take the CCL2 standard product and add it to the enzyme-labeled reaction plate respectively, and add 100 μl to each well as a standard well; the enzyme-labeled reaction plate is a sample well except the above-mentioned standard well, and add 50 μl of diluent and 50μl test sample;

[0066] (4) Add biotinylated antibody: Add biotin-labeled anti-human CCL2 antibody to the standard well and sample well of the enzyme-labeled reaction plate, a...

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Abstract

The invention provides a human serum CCL2 enzyme-linked immunosorbent assay kit as well as preparation and use methods thereof. The human serum CCL2 enzyme-linked immunosorbent assay kit comprises an enzyme-labeled reaction plate, a CCL2 standard product, biotin-labeled antihuman CCL2 antibodies and streptavidin-labeled horse radish peroxidase, wherein the enzyme-labeled reaction plate is coated with mouse-anti-human CCL2 monoclonal antibodies. The preparation method of the human serum CCL2 enzyme-linked immunosorbent assay kit comprises the steps of preparing the enzyme-labeled reaction plate, the CCL2 standard product, the biotin-labeled antihuman CCL2 antibodies and streptavidin-labeled horse radish peroxidase, wherein the enzyme-labeled reaction plate is coated with the mouse-anti-human CCL2 monoclonal antibodies. The use method of the human serum CCL2 enzyme-linked immunosorbent assay kit comprises the steps of adding samples, biotinylation antibodies and horse radish peroxidase, and carrying out color developing and terminating.

Description

technical field [0001] The invention relates to the field of immunological detection, in particular to a human serum CCL2 enzyme-linked immunological detection kit and a preparation and use method thereof. Background technique [0002] Human monocyte chemotactic protein-1 (CCL2 / Monocyte Chemotactic Protein-1, MCP-1), also known as monocyte chemoattractant and activator, is the main member of the CC subfamily in the chemokine superfamily, with Strong ability to induce and activate monocytes / macrophages. Cell types such as NK cells, T cells, eosinophils, basophils, etc. also respond to it. The cDNA of human monocyte chemoattractant encodes a precursor protein of 99 amino acid residues, and a hydrophobic signal peptide of 23 amino acid residues is cleaved to generate the final mature protein of 76 amino acid residues. Monocytes, mast cells, osteoblasts, fibroblasts, smooth muscle cells and many other cells can produce CCL2. [0003] CCL2 is a ligand for the seven-transmembra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68G01N2800/50G01N2800/7095
Inventor 张健卢奕李会强黄昕郭宏伟邹春林
Owner 广西南宁隆吉维特生物科技有限公司
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