Sequential detergent extraction prior to mass spectrometry analysis.

Fiona M. McCarthy; Amanda M. Cooksey; Shane C. Burgess

doi:10.1007/978-1-60327-310-7_7

Sequential detergent extraction prior to mass spectrometry analysis.

Fiona M. McCarthy, Amanda M. Cooksey, Shane C. Burgess

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

Sequential detergent extraction of proteins from eukaryotic cells has been used to increase proteome coverage of 2D-PAGE. We have adapted sequential detergent extraction for use with the high-throughput non-electrophoretic proteomics method of liquid chromatography and electrospray ionisation tandem mass spectrometry. This method of extraction yields comprehensive proteomes that include up to twice as many membrane proteins as other published methods. Two thirds of these membrane proteins have more than one transmembrane domain and many of these have multiple transmembrane domains. Since sequential detergent extraction (SDE) separates proteins based upon their physicochemistry and sub-cellular localisation, this method also provides useful data about cellular localisation.

Original language	English (US)
Pages (from-to)	110-118
Number of pages	9
Journal	Methods in molecular biology (Clifton, N.J.)
Volume	528
DOIs	https://doi.org/10.1007/978-1-60327-310-7_7
State	Published - 2009
Externally published	Yes

ASJC Scopus subject areas

Molecular Biology
Genetics

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10.1007/978-1-60327-310-7_7

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AB - Sequential detergent extraction of proteins from eukaryotic cells has been used to increase proteome coverage of 2D-PAGE. We have adapted sequential detergent extraction for use with the high-throughput non-electrophoretic proteomics method of liquid chromatography and electrospray ionisation tandem mass spectrometry. This method of extraction yields comprehensive proteomes that include up to twice as many membrane proteins as other published methods. Two thirds of these membrane proteins have more than one transmembrane domain and many of these have multiple transmembrane domains. Since sequential detergent extraction (SDE) separates proteins based upon their physicochemistry and sub-cellular localisation, this method also provides useful data about cellular localisation.

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Sequential detergent extraction prior to mass spectrometry analysis.

Abstract

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