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Detergent resistance as a tool in membrane research

Nature Protocols volume 2, pages 2159–2165 (2007)Cite this article

Abstract

The biological membrane is a complicated matrix wherein different lipid environments are thought to exist. The more ordered or raft environment has been perceived biochemically accessible via its relative resistance to detergent. This paper outlines the protocols developed in our laboratory for the analysis of such detergent-resistant membranes (DRMs). We stress the fact that DRMs are artifactual in nature and should not be equivocated to lipid rafts, their usefulness being limited to assigning raft-association potential most convincingly when changes in DRM composition are induced by biochemically/physiologically relevant events. These protocols are completed in 1–2 d.

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Figure 1: Triton X-100 (TX100) detergent-resistant membranes (DRMs) isolated from MDCK II cell homogenates using a linear sucrose gradient.
Figure 2: Triton X-100 (TX100) detergent-resistant membranes (DRMs) isolated from methyl-β-cyclodextrin treated (CD) and unmodified (−) MDCK II cell homogenates using a single-step OptiPrep gradient.

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Acknowledgements

We thank S. Schuck (University of California, San Francisco) for his helpful review of the manuscript. D.L. was the recipient of an international NSERC PGS scholarship. Work was supported by DFG SFB TR13 and DFG 1175 grants.

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  1. Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, Dresden, 01307, Germany

    Daniel Lingwood & Kai Simons

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Correspondence to Kai Simons.

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Lingwood, D., Simons, K. Detergent resistance as a tool in membrane research. Nat Protoc 2, 2159–2165 (2007). https://doi.org/10.1038/nprot.2007.294

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