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PMID: 7479734 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 92 ·No. 22 ·1995-10-24 ·Pages 10104-8

Smart EJ, Ying YS, Mineo C, Anderson RG

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.

MeSH Terms
Biomarkers Cell Fractionation/methods Cell Membrane/ultrastructure Cells, Cultured Electrophoresis, Polyacrylamide Gel Enzymes/analysis Fibroblasts/cytology,ultrastructure Humans Immunoblotting Microscopy, Electron Organelles/ultrastructure Proteins/analysis Skin/cytology,ultrastructure Ultracentrifugation
Chemicals
Biomarkers Enzymes Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Smart E J
Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical Center, Dallas 75235, USA.
Ying Y S
Mineo C
Anderson R G
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.92.22.10104
Published
1995-10-24
Pages
10104-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC40744
Subset
IM
Grants
NIGMS NIH HHS · GM 15631 · United States
NIGMS NIH HHS · GM 43169 · United States
NHLBI NIH HHS · HL 20948 · United States
External Links
PubMed source DOI: 10.1073/pnas.92.22.10104 PMC Full Text View journal details
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