Teacher
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TIMPERIO Anna Maria
(syllabus)
Introduction: "Homic" sciences and biology of complex systems. The revolution of the posterior hypothesis and the search for new hypotheses. What are the "omic" sciences ?. From proteins to proteomics, from metabolites to metabolomics and from lipids to lipidomics. Relationship between proteomics and other post-genomic sciences. Technology platforms: Sample preparation (plants, bacteria, animals): preparation of the protein extract for DIGE analysis, general guidelines for sample preparation for LC-MS analysis. Analysis of protein expression, metabolites and lipids by electrophoretic techniques Applications of the SDS-PAGE and the IEF to proteomics, metabolomics and lipidomics. DIfferential Gel Electrophoresis (DIGE), 2D-DIGE saturation Capillary electrophoresis (CE), two-dimensional electrophoresis (2DE). Applications of 2-DE in proteomics: limits and perspectives. Blue Native Electrophoresis on polyacrylamide gel (Blue Native PAGE). Clear Native PAGE (CN-PAGE). Gel free electrophoresis Electrophoresis on cellulose acetate Post analysis detection methods: Reverse coloring, Fluorescent dyes, Image analysis for multiplexing technologies Detection methods of post-translational modifications in mass spectrometry: Glycosylation, Phosphorylation, Nitration, Carbonylation Protein expression analysis by chromatographic techniques: Bottom-up proteomic approach Middle-down proteomic approach Top-down proteomic approach Multidimensional separations Mass spectrometry: introduction and concepts of resolution and accuracy The most used sources in proteomics: MALDI, ESI The most used sources in the metabolomics field: EI The most used sources in the lipidomic field: ESI, MALDI The analyzers: quadrupole, TOF, and ion traps, ORBITRAP, hybrids Tandem mass spectrometry (MS \ MS) Quantitative methods: reverse phase (RP) liquid chromatography (LC) systems coupled to mass spectrometry analysis. Relative quantification strategies based on marking with stable isotopes (iTRAQ, SILAC spike-in, Super-SILAC and organisms SILAC, ICAT, O18) Relative quantification strategies without marking ("label-free" methods: intensity-based and spectral-counting, calculation of PAI and emPAI). Targeted quantitative proteomics, metabolomics and lipidomics: SRM Coupled techniques: GC-MS, EC-MS, LC-MS \ MS, MudPIT Anna Maria Timperio
(reference books)
As a specific text is not yet on the market, the teacher will take care to report appropriate articles and magazines during the year
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