ocular proteomics with emphasis on two-dimensional gel electrophoresis and mass spectrometry眼蛋白质组学与强调二维凝胶电泳和质谱分析.pdfVIP

Ocular Proteomics with Emphasis on Two-Dimensional Gel Electrophoresis and Mass Spectrometry Nakul Mandal, Steffen Heegaard, Jan Ulrik Prause, Bent Honoré, and Henrik Vorum 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 polyacryl- amide gel electrophoresis (2D) 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, 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. Keywords: ocular proteomics, sample preparation, 2D, 2D-DIGE, mass spectrometry. Abbreviations 1D One-dimensional BME ß-mercaptoethanol 2D Two-dimensional 16-BAC Benzyldimethyl-n-hexadecylammonium chloride AEBSF 4-(2-Aminoethyl) benzenesulphonyl fluoride hydrochloride AMD Age-related macular degeneration ASB-14 Amidosulphobetaine-14 BCA Bicinchoninic acid CD Cluster of differentiation molecule Bent Honoré and Henrik Vorum share senior authorship. Shulin Li (ed.), Biological Proce