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Immunofluorescence kit for quantitatively detecting blood vessel endothelial marker CD105

A technology for quantitative detection of vascular endothelium, applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of long operation time, only qualitative, poor accuracy, etc., and achieve broad market prospects, reasonable design, and convenient use Effect

Pending Publication Date: 2018-12-28
NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the method for detecting CD105 is mainly enzyme-linked immunoassay (ELISA). ELISA technology has the following disadvantages: poor accuracy, long operation time, low degree of automation, low sensitivity, generally only qualitative, not quantitative, especially poor repeatability This shortcoming limits its clinical application, especially not suitable for the situation where accurate quantification is needed to help diagnose diseases. Therefore, the detection of CD105 by enzyme-linked immunosorbent technology is not suitable for rapid clinical diagnosis. How can a rapid quantitative detection be made? Equipment becomes an urgent problem

Method used

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  • Immunofluorescence kit for quantitatively detecting blood vessel endothelial marker CD105
  • Immunofluorescence kit for quantitatively detecting blood vessel endothelial marker CD105
  • Immunofluorescence kit for quantitatively detecting blood vessel endothelial marker CD105

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Experimental program
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Effect test

Embodiment 1

[0027] The structure of the immunofluorescence assay kit of the present invention;

[0028] Such as Figure 1-4 As shown, an immunofluorescence kit for quantitatively detecting the vascular endothelial marker CD105 includes a kit body and a test strip disposed in the kit body, the kit body includes a first box body, a second box body Body and the connecting strip connecting the first box body and the second box body. The first box body includes an upper shell and a lower shell. The upper shell and the lower shell are connected by clamping. Sample hole, observation window, one side of the observation window is provided with a detection mark, and the upper surface of the upper shell is also provided with a number of anti-skid protrusions, which are arranged in a rectangular array, and a number of anti-skid strips are provided on both sides of the lower end of the upper shell , a number of anti-slip strips are arranged equidistantly. There are clamping strips on the inner surfac...

Embodiment 2

[0037] The preparation method of the immunofluorescence assay kit of the present invention;

[0038] An immunofluorescence kit for quantitative detection of vascular endothelial marker CD105 comprises the following preparation steps:

[0039] (1) Preparation of FITC-labeled monoclonal antibody I: Dilute CD105 monoclonal antibody I with PBS buffer (0.05M, pH=7.2) containing 0.05mol / l NaCl at 4°C to make the final CD105 monoclonal antibody I The concentration is 10 mg / ml, add FITC fluorescein to the CD105 monoclonal antibody Ⅰ solution, the ratio of FITC fluorescein to CD105 monoclonal antibody Ⅰ is 1:50, stir at 4°C for 12 hours, add half-saturated ammonium sulfate, centrifuge, and Remove the precipitate, then dialyze with PBS buffer (0.01M, pH=7.2) to remove ammonium sulfate, add 0.01w / v% thimerosal to the prepared FITC-labeled antibody, divide into 1ml ampoules, and store at 4°C;

[0040] (2) Preparation of monoclonal antibody binding pad: Dilute the FITC-labeled CD105 monoc...

Embodiment 3

[0045] The preparation method of the immunofluorescence assay kit of the present invention;

[0046] An immunofluorescence kit for quantitative detection of vascular endothelial marker CD105 comprises the following preparation steps:

[0047] (1) Preparation of FITC-labeled monoclonal antibody I: Dilute CD105 monoclonal antibody I with PBS buffer (0.05M, pH=7.2) containing 0.15mol / l NaCl at 4°C to make the final CD105 monoclonal antibody I The concentration is 20 mg / ml, add FITC fluorescein to the CD105 monoclonal antibody Ⅰ solution, the ratio of FITC fluorescein to CD105 monoclonal antibody Ⅰ is 1:40, stir at 4°C for 16 hours, add half-saturated ammonium sulfate, centrifuge, and Remove the precipitate, then dialyze with PBS buffer (0.01M, pH=7.2) to remove ammonium sulfate, add 0.01w / v% thimerosal to the prepared FITC-labeled antibody, divide into 1ml ampoules, and store at 4°C;

[0048] (2) Preparation of monoclonal antibody binding pad: Dilute the FITC-labeled CD105 monoc...

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Abstract

The invention discloses an immunofluorescence kit for quantitatively detecting blood vessel endothelial markers CD105. The immunofluorescence kit comprises a kit body and a test strip arranged in thekit body; the kit comprises a second body, a trace blood-collecting vessel, a peripheral blood taking needle, a sucker, and an alcohol prep pad are arranged in the second body, full process of the blood drawing and blood test can be accomplished once; a detection line and a quality control line are arranged on the test strip; when being loaded on a sample pad, a to-be-detected sample moves forwardthrough capillary action, firstly reacts with a fluorescent mark antibody on a conjugate pad to form a fluorescent mark antibody-antigen compound; the to-be-tested sample continuously move forward, and combines with an antibody II cladded by the detection line to form the fluorescent mark antibody-antigen-antigen sandwich compound to immobilize on the detection line; the redundant fluorescent mark antibody and the rabbit anti-mouse IgG are combined and immobilized, and then detected through a fluorescence detector. The immunofluorescence kit disclosed by the invention has the advantages of being quantitative, fast, simple and convenient, and sensitive, and extensive in market prospect.

Description

technical field [0001] The invention relates to the field of in vitro diagnosis, in particular to an immunofluorescence kit for quantitatively detecting vascular endothelial marker CD105. Background technique [0002] CD105 is a marker of endothelial cell proliferation and one of the components of the transforming growth factor β (TGFβ) receptor complex. CD105 is overexpressed in proliferating endothelial cells and neovascular endothelial cells in tumor tissues with vigorous angiogenesis, while it is weakly or not expressed in vascular endothelial cells in normal mature tissues. It has the functions of regulating the response of endothelial cells to TGF, promoting endothelial cell proliferation and angiogenesis, etc. It can be used to diagnose tumors, predict curative effect, and can also be used for anti-tumor blood vessel therapy. [0003] At present, the method for detecting CD105 is mainly enzyme-linked immunoassay (ELISA). ELISA technology has the following disadvantag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/58G01N33/533
CPCG01N33/533G01N33/558G01N33/582G01N2333/71G01N2333/495
Inventor 唐静陈星星周鸿章
Owner NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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