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

Eric J. Smart, Yun Shu Ying, Chieko Mineo, Richard G W Anderson

Research output: Contribution to journalArticle

658 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)10104-10108
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume92
Issue number22
DOIs
StatePublished - Oct 24 1995

Fingerprint

Caveolae
Detergents
Cell Culture Techniques
Membranes
Octoxynol
Caveolins
Proteins

Keywords

  • epidermal growth factor receptor
  • folate receptor
  • G proteins

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

A detergent-free method for purifying caveolae membrane from tissue culture cells. / Smart, Eric J.; Ying, Yun Shu; Mineo, Chieko; Anderson, Richard G W.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 92, No. 22, 24.10.1995, p. 10104-10108.

Research output: Contribution to journalArticle

@article{b4ab152492c64e2ea111d46e59bc96b2,
title = "A detergent-free method for purifying caveolae membrane from tissue culture cells",
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.",
keywords = "epidermal growth factor receptor, folate receptor, G proteins",
author = "Smart, {Eric J.} and Ying, {Yun Shu} and Chieko Mineo and Anderson, {Richard G W}",
year = "1995",
month = "10",
day = "24",
doi = "10.1073/pnas.92.22.10104",
language = "English (US)",
volume = "92",
pages = "10104--10108",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "22",

}

TY - JOUR

T1 - A detergent-free method for purifying caveolae membrane from tissue culture cells

AU - Smart, Eric J.

AU - Ying, Yun Shu

AU - Mineo, Chieko

AU - Anderson, Richard G W

PY - 1995/10/24

Y1 - 1995/10/24

N2 - 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.

AB - 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.

KW - epidermal growth factor receptor

KW - folate receptor

KW - G proteins

UR - http://www.scopus.com/inward/record.url?scp=0028820041&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028820041&partnerID=8YFLogxK

U2 - 10.1073/pnas.92.22.10104

DO - 10.1073/pnas.92.22.10104

M3 - Article

VL - 92

SP - 10104

EP - 10108

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 22

ER -