Abstract
Lipid rafts were first defined as detergent-resistant membranes (DRMs) due to their relative insolubility in non-ionic detergents. Although they should not be confused with lipid rafts, DRMs are a valuable starting point for the study of these membrane domains and the interactions of proteins with rafts.
Here we describe the isolation of DRMs by ultracentrifugation on a sucrose gradient, a method we have used to study the role of lipid rafts in NKG2D-mediated signaling. We also describe raft fractionation of NK cells involving the selective solubility of β-octylglucoside (β-OG). OG is a non-ionic detergent that efficiently dissolves DRMs but does not disrupt protein associations with the cytoskeleton. Using these two techniques may yield useful information about the proteins involved in receptor recruitment into lipid rafts and the interactions of the actin cytoskeleton with lipid rafts.
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Acknowledgements
This work was supported by grants from the Spanish Fondo de Investigaciones Sanitarias-Fondos FEDER European Union (FISPI12/02587) from “Plan Nacional de I + D + I 2008–2011”, and the PCTI 2013–2017 (GRUPIN14-030) from Principado de Asturias, Spain.
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Serrano-Pertierra, E., López-Larrea, C. (2016). Using NK Cell Lipid Raft Fractionation to Understand the Role of Lipid Rafts in NK Cell Receptor Signaling. In: Somanchi, S. (eds) Natural Killer Cells. Methods in Molecular Biology, vol 1441. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3684-7_11
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DOI: https://doi.org/10.1007/978-1-4939-3684-7_11
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Publisher Name: Humana Press, New York, NY
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