An improved detergent-compatible gel-fractionation LC-LTQ-Orbitrap-MS workflow for plant and microbial proteomics

Luis Valledor; Wolfram Weckwerth

doi:10.1007/978-1-62703-631-3_25

An improved detergent-compatible gel-fractionation LC-LTQ-Orbitrap-MS workflow for plant and microbial proteomics

Luis Valledor
, Wolfram Weckwerth

Publications: Contribution to book › Chapter › Peer Reviewed

Abstract

In proteomics, liquid chromatography coupled to mass spectrometry (LC-MS/MS) is an invaluable technique to accurately identify and quantify large sets of proteins. In this chapter we show a time-effective, and detergent compatible, Ge-LC-LTQ-Orbitrap/MS proteomics workflow. The compatibility of this protocol with high concentrations of detergents significantly increases the extraction yield and the abundance of membrane proteins while gel fractionation increases the number of protein identifications. In our hands this workflow allows the identification of more than 1,500 proteins per sample, harvesting less than 20 mg of fresh weight, in many different organisms such as Chlamydomonas, Cyanothece, Arabidopsis, or Nicotiana, various microbes and enriched microbial samples.

Original language	English
Title of host publication	Plant Proteomics
Subtitle of host publication	Methods and Protocols - Methods in Molecular Biology
Editors	Jesus V. Jorrin-Novo, Setsuko Komatsu, Wolfram Weckwerth, Stefanie Wienkoop
Place of Publication	New York [u.a.]
Publisher	Humana Press
Pages	347-358
Number of pages	12
Volume	1072
ISBN (Electronic)	978-1-62703-631-3
ISBN (Print)	978-1-62703-630-6
DOIs	https://doi.org/10.1007/978-1-62703-631-3_25
Publication status	Published - 2014

Austrian Fields of Science 2012

106037 Proteomics
104002 Analytical chemistry

Keywords

Green proteomics
LC-MS
Orbitrap
Detergent-compatible

Access to Document

10.1007/978-1-62703-631-3_25

Cite this

Valledor, L., & Weckwerth, W. (2014). An improved detergent-compatible gel-fractionation LC-LTQ-Orbitrap-MS workflow for plant and microbial proteomics. In J. V. Jorrin-Novo, S. Komatsu, W. Weckwerth, & S. Wienkoop (Eds.), Plant Proteomics: Methods and Protocols - Methods in Molecular Biology (Vol. 1072, pp. 347-358). Humana Press. https://doi.org/10.1007/978-1-62703-631-3_25

Valledor, Luis ; Weckwerth, Wolfram. / An improved detergent-compatible gel-fractionation LC-LTQ-Orbitrap-MS workflow for plant and microbial proteomics. Plant Proteomics: Methods and Protocols - Methods in Molecular Biology. editor / Jesus V. Jorrin-Novo ; Setsuko Komatsu ; Wolfram Weckwerth ; Stefanie Wienkoop. Vol. 1072 New York [u.a.] : Humana Press, 2014. pp. 347-358

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Valledor, L & Weckwerth, W 2014, An improved detergent-compatible gel-fractionation LC-LTQ-Orbitrap-MS workflow for plant and microbial proteomics. in JV Jorrin-Novo, S Komatsu, W Weckwerth & S Wienkoop (eds), Plant Proteomics: Methods and Protocols - Methods in Molecular Biology. vol. 1072, Humana Press, New York [u.a.], pp. 347-358. https://doi.org/10.1007/978-1-62703-631-3_25

An improved detergent-compatible gel-fractionation LC-LTQ-Orbitrap-MS workflow for plant and microbial proteomics. / Valledor, Luis; Weckwerth, Wolfram.
Plant Proteomics: Methods and Protocols - Methods in Molecular Biology. ed. / Jesus V. Jorrin-Novo; Setsuko Komatsu; Wolfram Weckwerth; Stefanie Wienkoop. Vol. 1072 New York [u.a.]: Humana Press, 2014. p. 347-358.

Publications: Contribution to book › Chapter › Peer Reviewed

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AB - In proteomics, liquid chromatography coupled to mass spectrometry (LC-MS/MS) is an invaluable technique to accurately identify and quantify large sets of proteins. In this chapter we show a time-effective, and detergent compatible, Ge-LC-LTQ-Orbitrap/MS proteomics workflow. The compatibility of this protocol with high concentrations of detergents significantly increases the extraction yield and the abundance of membrane proteins while gel fractionation increases the number of protein identifications. In our hands this workflow allows the identification of more than 1,500 proteins per sample, harvesting less than 20 mg of fresh weight, in many different organisms such as Chlamydomonas, Cyanothece, Arabidopsis, or Nicotiana, various microbes and enriched microbial samples.

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KW - Detergent-compatible

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BT - Plant Proteomics

A2 - Jorrin-Novo, Jesus V.

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PB - Humana Press

CY - New York [u.a.]

ER -

An improved detergent-compatible gel-fractionation LC-LTQ-Orbitrap-MS workflow for plant and microbial proteomics

Abstract

Austrian Fields of Science 2012

Keywords

Access to Document

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