Learning outcomes of the course unit
The course aims at providing a systematic introduction to the main biochemical techniques for the identification, purification and functional characterization of macromolecules.
Pre-requisites (strongly recommended): basic courses in Physics, Chemistry, Microbiology and Biochemistry.
Course contents summary
Centrifugative techniques: differential centrifugation, density gradient centrifugation.
Protein extraction and purification.
Chromatography of biomacromolecules: ion-exchange, size exclusion and affinity chromatography. HPLC.
Electrophoresis: principles and applications. SDS-PAGE, isoelectrofocusing, 2D-PAGE.
Immunochemical techniques: immunoprecipitation, ELISA, Western blotting.
Spectrophotometry and fluorescence spectroscopy.
Use of radioisotopes in the biochemistry laboratory. Geiger counters, scintillation counters, autoradiography and phosphorimaging
Enzyme assays: continuous, discontinuous and based on coupled reactions. Activity units.
Introduction to mass spectrometry.
Rob Reed, David Holmes, Jonathan Weyers, Allan Jones
Metodologie di base per le scienze biomolecolari
The course is constituted by formal classroom presentations.
The final assessment of the students will be based on a written test.