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Bioscience Reports (2005) 25, (19–32) (Printed in Great Britain)

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Protein Detection Methods in Proteomics Research

Reiner WESTERMEIER^*1 and Rita MAROUGA^†

^*GE Healthcare Bio-Sciences, Munzinger Strasse 9, D-79111, Freiburg, Germany, ^†GE Healthcare Bio-Sciences, Björkgatan 30, S-751 82, Uppsala, Sweden

Abstract

In proteomics research chemical as well as physical methods are used to detect proteins subsequently to their separation. Physical methods are mostly applied after chromatography. They are either based on spectroscopy like light absorption at certain wavelengths or mass determination of peptides and their fragments with mass spectrometry. Chemical methods are used after two-dimensional electrophoresis and employ staining with organic dyes, metal chelates, fluorescent dyes, complexing with silver, or pre-labeling with fluorophores. In some cases autoradiography is still used. Since all of these techniques are very different in terms of sensitivity, their usefulness for quantitative determinations varies significantly. This review will describe the various protein detection methods applied to electrophoresis gels.

Key words: Blotting, protein staining, protein labeling, two-dimensional electrophoresis, quantification, DIGE

Abbreviations used: 2-D, two-dimensional; CCD, charge coupled device; DIGE, difference gel electrophoresis; EDTA, Ethylenediaminetetraacetic acid; ICAT, isotope coded affinity tag; MALDI ToF, matrix assisted laser desorption ionization time of flight; NMR, nuclear magnetic resonance; PTM, post-translational modification; PAS, periodic acid/Schiff's reagent; PVDF, Polyvinylidenefluoride; SDS, sodium dodecyl sulphate; Tris, Tris(hydroxymethyl)-aminoethane

¹To whom correspondence should be addressed (email: reiner.westermeier@ge.com)