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Application of UCP1 (uncoupling protein 1) to activity detection of human recombinant FGF21 (fibroblast growth factor 21) protein

A technology for FGF21 and protein activity, applied in the application field of UCP1 in the detection of human recombinant FGF21 protein activity, can solve the problems of high environmental requirements and high cost of detection methods, and achieve low environmental requirements, good practical value, and simple operation process Effect

Inactive Publication Date: 2017-10-10
XIAN MEDICAL UNIV
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Problems solved by technology

[0005] The purpose of the present invention is to provide an application of UCP1 in the detection of human recombinant FGF21 protein activity, which provides a new idea for judging the activity of human recombinant FGF21 protein, and solves the problems of high cost and high environmental requirements of existing detection methods

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  • Application of UCP1 (uncoupling protein 1) to activity detection of human recombinant FGF21 (fibroblast growth factor 21) protein
  • Application of UCP1 (uncoupling protein 1) to activity detection of human recombinant FGF21 (fibroblast growth factor 21) protein
  • Application of UCP1 (uncoupling protein 1) to activity detection of human recombinant FGF21 (fibroblast growth factor 21) protein

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Embodiment Construction

[0023] The application of the intraadipocyte uncoupling protein 1 (UCP1) group disclosed in the present invention in the detection of human recombinant FGF21 protein activity will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0024] 1. Cell culture and treatment

[0025] Mouse preadipocyte cells were planted in a 12-well culture plate and cultured in an incubator at a temperature of 37°C. Replace the fresh culture medium every day; when the cells grow to contact inhibition for 2 days, change to the induction medium, the induction medium is DMEM medium, which is added with 10% volume fraction of newborn bovine serum, 0.3IU / ml insulin, 20μmol / L Dexamethasone and 20 μmol / L rosiglitazone were treated for 2 days; replaced with insulin culture solution, the insulin culture solution was DMEM culture solution, in which 10% newborn bovine serum and 0.3IU / ml insulin were added, and the culture was continued After 2 days, change t...

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Abstract

The invention discloses an application of UCP1 (uncoupling protein 1) to activity detection of human recombinant FGF21 (fibroblast growth factor 21) protein. Adipocytes subjected to induced differentiation are taken as objects, qRT-PCR (quantitative real-time polymerase chain reaction) is adopted to detect the rising degree of UCP1 gene expression level of the adipocytes processed by the human recombinant FGF21 protein with different concentrations, and the activity of the human recombinant FGF21 protein is determined. The invention discovers that the UCP1 expression level and the concentration of the used human recombinant FGF21 protein have a good dose-effect relationship, so that the UCP1 expression level of the adipocytes is determined to be a new index for activity detection of the human recombinant FGF21 protein. A method adopts a simple operation process, has low environmental requirement and has good practical value.

Description

technical field [0001] The invention belongs to the technical field of uncoupling protein 1 (UCP1) application, and in particular relates to an application of UCP1 in the activity detection of human recombinant FGF21 protein. Background technique [0002] Drug development related to obesity and type 2 diabetes has always been a hot spot in the field of scientific research and development. Studies in obese and type 2 diabetes model mice and human patients have found that FGF21 recombinant protein can reduce body weight, lower blood lipids, lower blood sugar, significantly reduce obesity, type 2 diabetes and its complications. Therefore, among the many candidate molecules for drug development related to obesity and type 2 diabetes, fibroblast growth factor 21 (FGF21) is a research hotspot. [0003] Establishing a stable and efficient drug effect detection system is the key to evaluating candidate drugs. A detection index for objectively evaluating the therapeutic effect of F...

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6883C12Q2531/113C12Q2521/107
Inventor 赵玉峰陈镝梁向艳苏兴利魏兰兰赵妍妍谢荣刘英光
Owner XIAN MEDICAL UNIV
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