Proteome analysis using isoelectric focusing in immobilized pH gradient gels followed by mass spectrometry

Francesco Giorgianni, Dominic M. Desiderio, Sarka Beranova

Research output: Contribution to journalReview article

41 Citations (Scopus)

Abstract

Over the past several years, a large effort has been focused on improvements of two-dimensional (2-D) gel electrophoresis-based proteomics technology, and on development of novel approaches for proteome analysis. Here, we describe the application of an alternative strategy for the analysis of complex proteomes. The strategy combines isoelectric focusing in immobilized pH gradient strips (in-gel IEF), mass spectrometry (MS), and bioinformatics. A protein mixture is separated by in-gel IEF, and the entire strip is cut into a set of gel sections. Proteins in each gel section are digested with trypsin, and the tryptic peptides are subjected to liquid chromatography-nanoelectrospray-quadrupole ion-trap tandem mass spectrometry (LC-ESI-MS/MS). The LC-ESI-MS/MS data are used to identify the proteins through searches of a protein sequence database. Using this in-gel IEF-LC-MS/MS strategy, we have identified 127 proteins from a human pituitary. This study demonstrates the potential of the in-gel IEF-LC-MS/MS approach for analyses of complex mammalian proteomes.

Original languageEnglish (US)
Pages (from-to)253-259
Number of pages7
JournalElectrophoresis
Volume24
Issue number1-2
StatePublished - Jan 2003

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Proton-Motive Force
Isoelectric Focusing
Proteome
Mass spectrometry
Tandem Mass Spectrometry
Mass Spectrometry
Gels
Electrospray Ionization Mass Spectrometry
Proteins
Protein Databases
Electrophoresis, Gel, Two-Dimensional
Computational Biology
Liquid Chromatography
Proteomics
Trypsin
Ions
Technology
Liquid chromatography
Bioinformatics
Peptides

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry

Cite this

Proteome analysis using isoelectric focusing in immobilized pH gradient gels followed by mass spectrometry. / Giorgianni, Francesco; Desiderio, Dominic M.; Beranova, Sarka.

In: Electrophoresis, Vol. 24, No. 1-2, 01.2003, p. 253-259.

Research output: Contribution to journalReview article

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