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Determination kit for Golgi protein 73 and chemiluminiscence determination method of Golgi protein 73

A Golgi body and chemiluminescence technology, which is applied in chemiluminescence/bioluminescence, biological testing, and analysis through chemical reactions of materials, can solve problems such as poor stability, narrow detection range, and low sensitivity, and achieve shortened The effect of detection time, good stability and high sensitivity

Pending Publication Date: 2020-06-12
北京森美希克玛生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Enzyme-linked immunoassay technology is a traditional classic immunoassay technology, which has the advantages of simplicity, convenience, intuition and low cost, but also has some defects, such as low sensitivity, long reaction time, poor repeatability, narrow detection range and stability no problem

Method used

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  • Determination kit for Golgi protein 73 and chemiluminiscence determination method of Golgi protein 73
  • Determination kit for Golgi protein 73 and chemiluminiscence determination method of Golgi protein 73
  • Determination kit for Golgi protein 73 and chemiluminiscence determination method of Golgi protein 73

Examples

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Embodiment 1

[0054] This example provides a Golgi protein 73 assay kit, which includes a magnetic particle reagent, a GP73-binding antibody, a GP73-labeled antibody, a calibrator, a pre-excitation solution, an excitation solution, and a cleaning working solution.

[0055] The preparation method of the reagents in the assay kit is as follows.

[0056] (1) The preparation method of magnetic particle reagent:

[0057] The magnetic particles and the antibody of fluorescein isothiocyanate were mixed in 0.5M MES solution according to the ratio of 100:1, and equal amounts of EDC and NHS were added, and the reaction was carried out at room temperature for 60 min.

[0058] After magnetic separation, add 0.050M PBS containing 0.5% BSA to block for 1 h at room temperature, and then dilute to appropriate working concentration with 0.10M pH7.5 phosphate buffer containing 1% BSA to prepare a combined magnetic particle reagent and store at 4°C (2~8℃ can be used).

[0059] (2) The preparation method of ...

Embodiment 2

[0072] This embodiment provides a method for assaying Golgi protein 73, which includes using the assay kit provided in Embodiment 1 for detecting a sample to be tested, and specifically includes the following steps.

[0073] 50 μL of the test sample and calibrator were added to the reaction tube, and then incubated with the magnetic particle reagent and the GP73-binding antibody for 15 min at 37°C.

[0074] The incubated mixture was magnetically separated, the reaction solution was removed by suction, and the magnetic particles were washed three times with the cleaning working solution; then the washed mixture was incubated with GP73-labeled antibody at 37 °C for 15 min; then the reaction tube was placed Magnetic separation was carried out on the magnetic plate, the reaction solution was removed by suction, and the magnetic particles were washed 3 times with the cleaning working solution.

[0075] After washing, 100 μL of pre-excitation solution was added to the reaction tube,...

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Abstract

The invention discloses a determination kit for Golgi protein 73 and a chemiluminiscence determination method of the Golgi protein 73, and relates to the technical field of biology. Thedetection kit comprises a magnetic particle reagent, a GP73 binding antibody and a GP73 labeling antibody, combines a magnetic particle technology with an acridinium ester labeling technology, effectively shortens the detection time of Golgi protein 73, and is high in detection sensitivity, strong in specificity and good in stability.

Description

technical field [0001] The invention relates to the technical field of immunodetection, in particular, to a Golgi protein 73 assay kit and a chemiluminescence assay method thereof. Background technique [0002] GP73 (Golgi Protein73) is a type II transmembrane protein existing in the Golgi apparatus, with a molecular weight of 73KD. It has been confirmed that GP73 is a glycosylated protein, and N109 and N144 on its helical coil domain can undergo fucosylation. , glycosylation may be closely related to its function. The helical coil structure suggests that the protein can interact with other proteins through the extracellular region. [0003] Kladney study in 2000 showed that in normal human liver tissue, GP73 is mainly expressed by bile duct epithelial cells, while hepatocytes express little or no expression. GP73 expression can be up-regulated after hepatocytes are infected by virus. In 2005, Block et al. found that the level of GP73 was significantly increased in the ser...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N21/76
CPCG01N33/68G01N33/54326G01N21/76
Inventor 肖文峰胡国茂王国建
Owner 北京森美希克玛生物科技有限公司
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