Teacher
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TIMPERIO Anna Maria
(syllabus)
OMIC applied sciences Teacher: Prof. Anna Maria Timperio Definition of "omic" sciences, Description of the program and examination procedures. Safety standards in scientific laboratories. Proteomics definition: Expression and functional proteomics, proteomic analysis strategies. Native electrophoresis (BLUE-NATIVE gel), one and two-dimensional (1D-GEL, 2D-GEL). Iso-electro-focus (IEF) Reverse phase chromatography RP-HPLC. Proteomics applications for the study of cancer cells and for early diagnosis for the presence of biomarkers (examples) Laboratory 1, Protein extraction from material to be defined. Determination of protein concentration (Brendford method). Use of spectrophotometer Laboratory 2. One dimensional electrophoresis (1D-SDS-PAGE). Cutting of the bands from the gel and digestion in trypsin (without label) Laboratory 3. Reverse phase HPLC: construction of a gradient. Determination of the slope and study of the chromatogram Metabolomics definition: Sample preparation techniques for metabolome analysis.Chromatographic and mass spectrometry methods used to characterize metabolites that function as biomarkers Examples of metabolomics studies in clinical research: Application of metabolomics in the study of autism Lipidomics definition of main lipids and their determination in mass spectrometry, determination of lipids of red blood cell membranes Foodomica definition. determination of flavonoids, vitamins in food -Laboratory 4. Extraction of metabolites according to the (Bligh & Dyer) method Mass Spectrometry: General. Main sources, analyzers and detectors. Analysis of proteins, peptides, metabolites, lipids in intact mass. Tandem mass: meaning and purposes - Laboratory 5. Determination of the intact mass of a protein. Determination of the amino acid sequence with mass spectrometry (nanoESI-TRAP), Determination of the main lipid classes in ORBITRAP spectrometry, Determination of flavonoids in MALDI TOF spectrometry Complexity reduction: Subcellular fractionation. Lysis of cells and homogenization of tissues. Separation of subcellular fractions. Proteomics, metabolomics and lipidomics of the membrane Proteomics, metabolomics, lipidomics of organelles and intracellular compartments. Immunoprecipitation Immunoprecipitation to reduce the complexity of a sample for mass spectrometry. Functional Proteomics and Immunoprecipitation
(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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