A detergent-free method for purifying caveolae membrane from tissue culture cells

Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10104-8. doi: 10.1073/pnas.92.22.10104.

Abstract

Current methods for purifying caveolae from tissue culture cells take advantage of the Triton X-100 insolubility of this membrane domain. To circumvent the use of detergents, we have developed a method that depends upon the unique buoyant density of caveolae membrane. The caveolae fractions that we obtain are highly enriched in caveolin. As a consequence we are able to identify caveolae-associated proteins that had previously gone undetected. Moreover, resident caveolae proteins that are soluble in Triton X-100 are retained during the isolation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Biomarkers
  • Cell Fractionation / methods
  • Cell Membrane / ultrastructure*
  • Cells, Cultured
  • Electrophoresis, Polyacrylamide Gel
  • Enzymes / analysis
  • Fibroblasts / cytology
  • Fibroblasts / ultrastructure
  • Humans
  • Immunoblotting
  • Microscopy, Electron
  • Organelles / ultrastructure*
  • Proteins / analysis
  • Skin / cytology
  • Skin / ultrastructure*
  • Ultracentrifugation

Substances

  • Biomarkers
  • Enzymes
  • Proteins