Proteins of rat serum: I. Establishing a reference two-dimensional electrophoresis map by immunodetection and microbore high performance liquid chromatography-electrospray mass spectrometry

Paul Haynes, Ingrid Miller, Ruedi Aebersold, Manfred Gemeiner, Ivano Eberini, Maria Rosa Lovati, Cristina Manzoni, Mara Vignati, Elisabetta Gianazza

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58 Citations (Scopus)

Abstract

In the present investigation, we have identified 56 major spots, or spot rows, corresponding to 22 proteins, in the 2-DE pattern of adult male rats. This was done mainly by applying two complementary techniques, namely immunoblotting and high performance liquid chromatography-mass spectrometry (HPLC-MS) peptide mapping. Glycoproteins were characterized by affinity blotting with six lectins. We have also detailed how rat serum differs from human serum in two main respects: (i) relative abundance of individual proteins, which amounts in some cases to a complete absence in either sample, and (ii) varying molecular parameters for homologous proteins. It was thus possible to establish a first-generation reference map of rat serum proteins, which can be accessed through http://weber.u.washington.edu/∼ruedilab/aebersold.html. We hope the present database will be a useful reference for the evaluation of changes in serum protein distribution in the course of pharmacological and toxicological studies. The recognition of species-specific proteins appears of special relevance in this respect.
Original languageEnglish
Pages (from-to)1484-1492
Number of pages9
JournalElectrophoresis
Volume19
Issue number8-9
DOIs
Publication statusPublished - 1998
Externally publishedYes

Keywords

  • rat serum
  • two-dimensional polyacrylamide gel electrophoresis
  • electrospray mass spectrometry
  • microbore liquid chromatography

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