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

Author(s)
Luis Valledor, Wolfram Weckwerth
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.

Organisation(s)
Volume
1072
Pages
347-358
No. of pages
12
DOI
https://doi.org/10.1007/978-1-62703-631-3_25
Publication date
2014
Peer reviewed
Yes
Austrian Fields of Science 2012
106037 Proteomics, 104002 Analytical chemistry
Keywords
ASJC Scopus subject areas
Genetics, Molecular Biology
Portal url
https://ucris.univie.ac.at/portal/en/publications/an-improved-detergentcompatible-gelfractionation-lcltqorbitrapms-workflow-for-plant-and-microbial-proteomics(813df139-ec4f-433b-93bc-1849a0a428f3).html