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Isolation of renal proximal tubular brush-border membranes

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Abstract

This protocol describes a method for the isolation and purification of renal proximal tubular brush-border membranes in high yield and high purity. Based on a different reactivity of the brush-border membrane compared to other cellular membranes with divalent cations, such as Mg2+, purified membrane vesicles can be obtained after a few differential centrifugation steps (within approximately 3 h) that are suitable for in vitro studies, such as transport experiments or protein and lipid analysis.

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Figure 1: Electron microscopic picture of freeze-fractured purified rat renal BBMVs.

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Acknowledgements

Work related to this method has been continuously supported by the Swiss National Fonds.

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Authors and Affiliations

  1. Institute of Physiology, University Zürich, Winterthurerstrasse 190, Zürich, 8057, Switzerland

    Jürg Biber, Gerti Stange & Heini Murer

  2. Division of Clinical Pharmacology and Toxicology, Department of Medicine, University Hospital, Zurich, 8091, Switzerland

    Bruno Stieger

Authors
  1. Jürg Biber
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  2. Bruno Stieger
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  3. Gerti Stange
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  4. Heini Murer
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Correspondence to Jürg Biber.

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The authors declare no competing financial interests.

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Biber, J., Stieger, B., Stange, G. et al. Isolation of renal proximal tubular brush-border membranes. Nat Protoc 2, 1356–1359 (2007). https://doi.org/10.1038/nprot.2007.156

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