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

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 furthe...

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Detalles Bibliográficos
Autores principales: Lindner, Karina, van Ek, Larissa C., Awwad, Khader, Hanke, Franziska, Korepanova, Alla V., Rybin, Vladimir, Beckenbauer, Katharina, Gavin, Anne-Claude
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9722783/
https://www.ncbi.nlm.nih.gov/pubmed/36595894
http://dx.doi.org/10.1016/j.xpro.2022.101894
Descripción
Sumario: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)

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