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Course organization
Advanced proteomic: a view of concepts and technologies for the studying the proteome, including the latests developments.
1. Introduction. Definitions and types of proteomic analysis.
2. Chromatographic methods (bi and multi dimensional) applied to proteome analysis. Multidimensional protein idenfication technology (MudPIT) and MultiDimensional Liquid Chromatography (MDLC).
3. Electrophoretic methods for proteome analysis. Isoelectrofocusing (IEF) and SDS electrophoresis (SDS-PAGE). 2D Maps.
4. Sample preparation and handling for proteomic analysis.
5. Prefractionation of the sample for the proteome analysis of low abundance proteins.
6. Image analysis: PDQuest and other softwares.
7. Mass spectrometry analysis.
8. Tandem mass spectrometry MS/MS (Q-Tof, TripleQ, TOF/TOF, FT-ICR, IonTrap e MALDI PSD MS).
9. Searching engines for Data Base searching and Data Bases for protein identification.
10. Quantitative proteomic analysis: Fluorescent Dyes (DIGE); metabolic isotopic labelling (SILAC), chemical (ICAT, iTRAQ, AQUA, acrylamide d0/d3), enzimatical (H218O/ H216O); label free quantification (SELDI-MS).
11. Post-translational modifications characterization (IMAC; immunoprecipitation; biotin-switch).
12. Applications of proteomic analysis.
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