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Biol Proced Online. 2009 Dec 24;12(1):56-88. doi: 10.1007/s12575-009-9019-7.

Ocular proteomics with emphasis on two-dimensional gel electrophoresis and mass spectrometry.

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1
Eye Pathology Section, Department of Neuroscience and Pharmacology, University of Copenhagen, Copenhagen, Denmark.
2
Department of Medical Biochemistry, Aarhus University, Aarhus, Denmark.
3
Department of Ophthalmology, Aalborg Hospital, Aarhus University Hospital, Aalborg, Denmark.
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Contributed equally

Abstract

The intention of this review is to provide an overview of current methodologies employed in the rapidly developing field of ocular proteomics with emphasis on sample preparation, two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry (MS). Appropriate sample preparation for the diverse range of cells and tissues of the eye is essential to ensure reliable results. Current methods of protein staining for 2D-PAGE, protein labelling for two-dimensional difference gel electrophoresis, gel-based expression analysis and protein identification by MS are summarised. The uses of gel-free MS-based strategies (MuDPIT, iTRAQ, ICAT and SILAC) are also discussed. Proteomic technologies promise to shed new light onto ocular disease processes that could lead to the discovery of strong novel biomarkers and therapeutic targets useful in many ophthalmic conditions.

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