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Analytical and Bioanalytical Chemistry

, Volume 400, Issue 2, pp 415–421 | Cite as

Combination of acid labile detergent and C18 Empore™ disks for improved identification and sequence coverage of in-gel digested proteins

  • Henning Koehn
  • Benjamin Lau
  • Stefan Clerens
  • Jeffrey E. Plowman
  • Jolon M. Dyer
  • Umi Salamah Ramli
  • Santanu Deb-ChoudhuryEmail author
Technical Note

Abstract

A protocol for improved extraction of peptides from in-gel protein digests, using a combination of the acid labile surfactant, sodium deoxycholate (SDC) and C18 Empore™ membranes, is presented. This approach results in better mass spectrum quality, higher numbers of identified peptide peaks and improved identification scores compared to standard tryptic digestion protocols, or protocols using only SDC or only C18 Empore™ disks. The advantages of the new protocol are demonstrated for two different types of samples: Merino wool intermediate filament proteins and Elaeis guineensis (oil palm) mesocarp proteins.

Keywords

2-D electrophoresis MALDI-TOF In-gel digestion 

Notes

Acknowledgements

The authors wish to thank Mrs. Anita Grosvenor for editing this manuscript. Funding for this project was provided by the Foundation for Research Science and Technology (C10X0710).

Supplementary material

216_2011_4765_MOESM1_ESM.pdf (1 mb)
Electronic supplementary material (PDF 1025 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Henning Koehn
    • 1
  • Benjamin Lau
    • 2
  • Stefan Clerens
    • 1
  • Jeffrey E. Plowman
    • 1
  • Jolon M. Dyer
    • 1
  • Umi Salamah Ramli
    • 2
  • Santanu Deb-Choudhury
    • 1
    Email author
  1. 1.AgResearch LtdChristchurchNew Zealand
  2. 2.Malaysian Palm Oil BoardAdvanced Biotechnology and Breeding CentreKajangMalaysia

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