In vitro and in cellulo ApoE particle formation, isolation, and characterization

Summary

Apolipoprotein E (ApoE) particles are responsible for packing and transporting lipids throughout aqueous environments. We detail steps to assess in vitro particles forming from artificial membranes using right-angle light scattering and to measure their size using dynamic light scattering. We further describe how to generate in cellulo ApoE particles containing triacylglycerol under fatty-acid-induced stress. We also detail steps to isolate them from cell secretome by immunoprecipitation and analyze their lipid cargo by thin-layer chromatography.

For complete details on the use and execution of this protocol, please refer to Lindner et al. (2022). ¹

Graphical abstract

Highlights

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  Collection of protocols that enable the analysis of ApoE lipoprotein particles

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  Characterization of ApoE particles generated in vitro using light scattering

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  ApoE lipidation in lipid droplet-loaded cultured cells

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  Isolation of particles and characterization of lipid cargo by thin layer chromatography

Abstract

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Abstract

Apolipoprotein E (ApoE) particles are responsible for packing and transporting lipids throughout aqueous environments. We detail steps to assess in vitro particles forming from artificial membranes using right-angle light scattering and to measure their size using dynamic light scattering. We further describe how to generate in cellulo ApoE particles containing triacylglycerol under fatty-acid-induced stress. We also detail steps to isolate them from cell secretome by immunoprecipitation and analyze their lipid cargo by thin-layer chromatography.