Assessment of Caveolae/Lipid Rafts in Isolated Cells

Methods Mol Biol. 2017:1527:251-269. doi: 10.1007/978-1-4939-6625-7_20.

Abstract

This chapter outlines protocols to evaluate protein localization, recruitment or phosphorylation levels in cholesterol/sphingolipids-enriched cell membrane domains and recommends experimental designs with pharmacological tolls to evaluate potential cell functions associated with these domains. We emphasize the need for the combination of several approaches towards understanding the protein components and cellular functions attributed to these distinct microdomains.

Keywords: Caveolae; Detergent-free method; Lipid rafts; Methyl-β-cyclodextrin; Statins.

MeSH terms

  • Animals
  • Carbonates / metabolism
  • Caveolae / metabolism*
  • Cell Membrane / metabolism
  • Cholesterol / chemistry
  • Cholesterol / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Hydrogen-Ion Concentration
  • Membrane Microdomains / chemistry
  • Membrane Microdomains / metabolism*
  • Muscle, Smooth, Vascular / cytology
  • Myocytes, Smooth Muscle / metabolism
  • Rats
  • Sphingolipids / chemistry
  • Sphingolipids / metabolism
  • beta-Cyclodextrins / chemistry
  • beta-Cyclodextrins / metabolism

Substances

  • Carbonates
  • Sphingolipids
  • beta-Cyclodextrins
  • methyl-beta-cyclodextrin
  • sodium carbonate
  • Cholesterol