A Detergent-Free Method for Purifying Caveolae Membrane from Tissue Culture Cells

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...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1995-10, Vol.92 (22), p.10104-10108
Main Authors: Smart, Eric J., Ying, Yun-Shu, Mineo, Chieko, Richard G. W. Anderson
Format: Article
Language:English
Subjects:
Antibodies
Biomarkers
Caveolae
Caveolins
Cell Fractionation - methods
Cell Membrane - ultrastructure
Cell membranes
Cells
Cells, Cultured
Cellular biology
Centrifugation
Cleaning
Cultured cells
Detergents
Electrophoresis, Polyacrylamide Gel
Endothelial cells
Enzymes - analysis
Fibroblasts - cytology
Fibroblasts - ultrastructure
Humans
Immunoblotting
Membranes
Microscopy, Electron
Organelles - ultrastructure
P branes
Proteins - analysis
Receptors
Skin - cytology
Skin - ultrastructure
Ultracentrifugation
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Summary: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.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.92.22.10104