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A method and kit for detecting the biological activity of apoa5 for non-diagnostic purposes

A technology of biological activity and kit, which is applied in the field of detection of biological activity of ApoA5, ​​and can solve problems such as elevated liver TG

Inactive Publication Date: 2016-06-29
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the research on the regulating effect of ApoA5 on TG in organisms is mainly focused on its ability to reduce TG in blood and fat cells, and at the same time increase TG in the liver. No other cells have been found to be used for ApoA5 to regulate TG. role research

Method used

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  • A method and kit for detecting the biological activity of apoa5 for non-diagnostic purposes
  • A method and kit for detecting the biological activity of apoa5 for non-diagnostic purposes
  • A method and kit for detecting the biological activity of apoa5 for non-diagnostic purposes

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The detection of embodiment 1 ApoA5 biological activity

[0037] Preparation of samples to be tested: the test is divided into a test group and a control group, and each group is treated as follows:

[0038] (1) Treatment of the test group: the HL-1 cardiomyocytes (mouse atrial myocytes) inoculated on a 12-well culture plate were mixed with 1×10 6 / cm 2 After the cells reached 90% confluence, they were starved for 12 hours with claycomb medium containing 1% serum, and divided into 3 treatments. 0.5mmol / L oleic acid, 1800ng / mL ApoA5+0.5mmol / L oleic acid in 1% serum claycomb medium was incubated for 24 hours.

[0039] (2) Treatment of the control group: the HL-1 cardiomyocytes (mouse atrial myocytes) inoculated on a 12-well culture plate were mixed with 1×10 6 / cm 2 After the cells reached 90% confluency, they were starved for 12 hours with 1% serum claycomb medium, and then added with 1% serum claycomb medium containing 0.5 mmol / L oleic acid for 24 hours.

[0040] D...

Embodiment 2

[0050] The detection of embodiment 2ApoA5 biological activity

[0051] Preparation of samples to be tested: the test is divided into a test group and a control group, and each group is treated as follows:

[0052] (1) Treatment of the test group: the HL-1 cardiomyocytes (mouse atrial myocytes) inoculated on a 12-well culture plate were mixed with 1×10 6 / cm 2 After the cells reached 90% confluence, they were starved for 12 hours with claycomb medium containing 1% serum, and divided into 3 treatments. 0.5mmol / L oleic acid, 1800ng / mL ApoA5+0.5mmol / L oleic acid in 1% serum claycomb medium was incubated for 24 hours.

[0053] (2) Treatment of the control group: the HL-1 cardiomyocytes (mouse atrial myocytes) inoculated on a 12-well culture plate were mixed with 1×10 6 / cm 2 After the cells reached 90% confluency, they were starved for 12 hours with 1% serum claycomb medium, and then added with 1% serum claycomb medium containing 0.5 mmol / L oleic acid for 24 hours.

[0054] Di...

Embodiment 3

[0059] The detection of embodiment 3ApoA5 biological activity

[0060] Preparation of samples to be tested: the test is divided into a test group and a control group, and each group is treated as follows:

[0061] (1) Treatment of the test group: the HL-1 cardiomyocytes (mouse atrial myocytes) inoculated on a 12-well culture plate were mixed with 1×10 6 / cm 2 After the cells reached 90% confluence, they were starved for 12 hours with claycomb medium containing 1% serum, and divided into 3 treatments. 0.5mmol / L oleic acid, 1800ng / mL ApoA5+0.5mmol / L oleic acid in 1% serum claycomb medium was incubated for 24 hours. The ApoA5 used has been inactivated and has lost its biological activity.

[0062] (2) Treatment of the control group: the HL-1 cardiomyocytes (mouse atrial myocytes) inoculated on a 12-well culture plate were mixed with 1×10 6 / cm 2 After the cells reached 90% confluency, they were starved for 12 hours with 1% serum claycomb medium, and then added with 1% serum ...

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Abstract

The invention relates to the field of protein activity detection, and in particular relates to a method and a kit for non-diagnostically detecting ApoA5 biological activity. The method comprises the following steps: mixing starved HL-1 myocardial cells with ApoA5 and oleic acid, incubating, splitting the cells, releasing TG and detecting to obtain a first TG concentration; mixing the starved HL-1 myocardial cells with oleic acid, incubating, splitting the cells, releasing TG and detecting to obtain a second TG concentration; comparing the first TG concentration with the second TG concentration to obtain the ApoA5 biological activity. The detection method provided by the invention can be used for correctly detecting whether the ApoA5 has biological activity or not.

Description

technical field [0001] The invention relates to the field of protein activity detection, in particular to a method and kit for non-diagnostic detection of ApoA5 biological activity. Background technique [0002] With the improvement of modern living standards and changes in lifestyles, the incidence of obesity is increasing. At the same time, the increase in lipids and ectopic deposition associated with obesity makes the incidence of obesity-related diseases increase year by year. Therefore, effectively reducing the lipid deposition associated with obesity is an important challenge facing medicine today. [0003] Obesity is the result of excessive storage of triglyceride (TG) in adipocytes due to the imbalance between energy intake and energy consumption, and the result of abnormal increase in cell volume. The root cause is abnormal lipid metabolism. The essence of abnormal lipid metabolism refers to the abnormality of lipoprotein and apolipoprotein. The activity of key en...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/61C12Q1/44C12Q1/02
Inventor 赵水平罗俊郑小燕聂赛赵旺
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV