Enzyme-linked immunosorbent assay kit and preparation method thereof

A preparation method and ELISA plate technology, applied in the field of biomedicine, can solve the problems of lack of FN1 research and the production of a complete set of FN1ELISA kits.

Active Publication Date: 2013-12-25
WUHAN SANYING BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The literature has widely reported the role of FN1 as a biomarker in the diagnosis and treatment of diseases, but currently, several major ELISA kit manufacturers in the world only produc

Method used

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  • Enzyme-linked immunosorbent assay kit and preparation method thereof
  • Enzyme-linked immunosorbent assay kit and preparation method thereof
  • Enzyme-linked immunosorbent assay kit and preparation method thereof

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Experimental program
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Embodiment Construction

[0051] 1. Reagents:

[0052] (1) Coating solution (0.02mol / l, sodium carbonate-sodium bicarbonate buffer at pH 9.6): Na 2 CO 3 0.6g, NaHCO 3 1.16g, Na 2 N 3 0.2g, add double distilled water to 1000ml, adjust pH to 9.6.

[0053] (2) Specimen diluent (0.01mol / l PBS containing 8% calf serum pH7.4): NaCl 8.0g, NaH 2 PO 4 2H 2 O 0.3g, Na 2 HPO 4 12H 2 O 2.9g, KCl 0.2g, thimerosal 0.1g, add double distilled water to 1000ml, adjust pH to 7.4.

[0054] (3) Blocking solution (8% calf serum / PBS solution): 80ml calf serum, 0.01mol / l pH7.4 PBS 920ml.

[0055] (4) Washing solution (pH7.4 PBST): NaCl 8.0g, NaH 2 PO 4 2H 2 O 0.3g, Na 2 HPO 4 12H 2 O 2.9g, Tween 20 0.5g, thimerosal 0.1g, add double distilled water to 1000ml, adjust pH to 7.4.

[0056] (5) Enzyme-labeled secondary antibody (HRP-labeled goat anti-rabbit polyclonal antibody): purchased from R&D systems in the United States, and diluted to 1:10000 with PBS before use.

[0057] (6) FN1 protein standard substa...

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Abstract

The invention relates to an enzyme-linked immunosorbent assay (ELISA) kit and a preparation method thereof, and more specifically relates to a novel anti-human FN1 protein monoclonal antibody coated ELISA plate and a preparation method of the ELISA kit. A key step of the preparation method is that: the specific mouse anti-human FN1 protein monoclonal antibody is prepared by genetic engineering method, and is purified; and then the ELISA plate is coated with the specific mouse anti-human FN1 protein monoclonal antibody, and is used as a constituent part of the ELISA kit. The ELISA kit also comprises FN1 recombined protein standard, polyclonal antibody of anti-human FN1, horse radish peroxidase-labeled second antibody, a sample diluent, a washing liquid, an antibody diluent, a colour reagent, and a stop solution. The ELISA kit can be used for rapid and convenient detection of FN1 concentration of human serum and blood plasma, and is mainly provided for scientific research or clinical research of cancer related diseases.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to an enzyme-linked immunoassay kit for detecting human fibronectin (FN1), and also relates to a preparation method of the kit. Background technique [0002] FN1 (Fibronectin 1) is type I fibronectin, a glycoprotein that exists in body fluids in a soluble form or in the extracellular matrix in an insoluble form, respectively. Plasma-soluble fibronectin is the main protein component in plasma. Insoluble cellular fibronectin is a major component of the extracellular matrix. Fibronectin is involved in cellular adhesion and migration processes, including embryonic development, wound healing, blood coagulation, host defense and metastasis. Plays an important role in cell adhesion, growth, migration and differentiation, and is important for processes such as wound healing and embryonic development. (Roumen, P. et al. Fibronectin at a glance[J]. J Cell Sci, 2002, 115, 386...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543
Inventor 赵以睿王慕元张鹏连绘
Owner WUHAN SANYING BIOTECH
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