Method for analyzing cholesterol outflow capacity promoted by HDL of different sizes, and application
A cholesterol and capacity technology, applied in analytical materials, biomaterial analysis, preparation of test samples, etc., can solve problems such as HDL differentiation
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Embodiment 1
[0048] Example 1 Determination of different HDL-promoted total cholesterol efflux rates in the plasma of obese patients
[0049] Such as figure 1 As shown, the mouse macrophage J774 cell line was inoculated in the wells of the culture plate and incubated, and then all were treated with 3 H isotope-labeled cholesterol medium to label the cells in the wells of the culture plate, and then incubate at 37°C for 16 hours without adding cAMP to induce the expression of total cholesterol transporters; take plasma from obese patients, and label as Obese, its plasma is separated by FPLC chromatography to separate HDL molecules of gradient size, such as figure 2 As shown, the components at 32, 34, and 37 are respectively selected to represent three HDL components with different particle sizes. It should be noted that 32, 34, and 37 are figure 2 The labels on the middle abscissa, the larger the value, the smaller the particles of HDL molecules in this group; add the three groups into ...
Embodiment 2
[0050] Example 2 Determination of different HDL-promoted total cholesterol efflux rates in plasma of healthy people
[0051] The mouse macrophage J774 cell line was inoculated in the wells of the culture plate and incubated, and then all were treated with 3 H isotope-labeled cholesterol culture solution labels the cells in the wells of the culture plate, and then incubates at 37°C for 16 hours without adding cAMP to induce the expression of total cholesterol transporters; take the plasma from healthy people and mark it as Healthy control, its plasma was separated by FPLC chromatography to separate HDL molecules of gradient size, such as figure 2 As shown, select the components at 32, 34, and 37 respectively to obtain three HDL components with different particle sizes; add the three groups to the wells of the cell culture plate, and incubate at 37°C for 4 hours, and then pipette The supernatant culture solution in the wells of the culture plate is centrifuged to separate the ...
Embodiment 3
[0052] Example 3 Determination of different HDL-based cholesterol efflux rates based on ABCA1 in the plasma of obese patients
[0053] The mouse macrophage J774 cell line was inoculated in the wells of the culture plate and incubated, and then all were treated with 3 Cells were labeled with H isotope-labeled cholesterol culture medium, then cAMP was added to the cells, and incubated at 37°C for 16 hours to induce ABCA1 expression; the plasma of obese patients was collected, labeled as Obese, and the plasma was separated by FPLC chromatography Gradient-sized HDL particles such as figure 2 As shown, select the components at 32, 34, and 37 respectively to obtain three HDL components with different particle sizes; add the three groups to the wells of the cell culture plate, and incubate at 37°C for 4 hours, and then pipette The supernatant culture solution in the wells of the culture plate is centrifuged to separate the residual cells in the supernatant culture solution, and the...
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