Università degli Studi di Perugia

Unit BIOCHEMICAL METHODS

Course
Biology
Study-unit Code
GP004082
Curriculum
Biosanitario
Teacher
Serena Porcellati
Teachers
  • Serena Porcellati
  • Sandra Buratta (Codocenza)
  • Elisabetta Chiaradia (Codocenza)
Hours
  • 25 ore - Serena Porcellati
  • 12 ore (Codocenza) - Sandra Buratta
  • 10 ore (Codocenza) - Elisabetta Chiaradia
CFU
6
Course Regulation
Coorte 2021
Offered
2021/22
Learning activities
Affine/integrativa
Area
Attività formative affini o integrative
Sector
BIO/10
Type of study-unit
Opzionale (Optional)
Type of learning activities
Attività formativa monodisciplinare
Language of instruction
Italian
Contents
Spectrophotometric techniques. Electrophoretic techniques. Mass spectrometry applied to the study of proteins
Reference texts
Keith Wilson and John Walker: Biochimica e biologia molecolare:Principi e tecniche¿ EDs. 2019 Raffaello Cortina Editore. ¿
M.Maccarrone: Metodologie Biochimiche e Biomolecolari, Zanichelli Editore
Bonaccorsi, Contestabile, Di Salvo -Metodologie biochimiche-Casa Editrice Ambrosiana. Distrib. Zanichell
Educational objectives
Knowledge about the spectrophotometric and electrophoretic methods for the study of the main biomolecules, as well as about the mass spectrometry methods for the identification of proteins
Skills: the student must be able to set up a protocol for the electroforesis and spectrophotometric qualitative/quantitative analysis of the main biomolecules as well as for the identification of specific proteins by mass spectrometry
Prerequisites
Basic knowledge in the field of organic chemistry and biochemistry
Teaching methods
Lectures with the help of slides
Other information
Optional
Learning verification modality
Interview on course topics , within the exam with the other teachers.
Extended program
Spectrophotometric techniques: introduction, ultraviolet and visible light spectroscopy, Lambert –beer Law and application. Spectrofluorimetry (2.5 hours).
Electrophoresis: the basics. Horizontal and Vertical Electrophoresis. SDS-PAGE and native electrophoresis. Gel staining: Comassie brilliant Blue, Silver staining, fluorescent dyes and usage criteria; Ethidium bromide. (2.5 hours).
Blotting Techniques: the basics. Southern and Northern Blotting, Western Blotting: Transfer, Protein Detection: by Membrane Dyeing (Ponceau S) or Enzymatic Conjugated Antibodies. Immunoblotting: the different phases (2.5 hours).
Proteomics: basics, gel based and gel free approach. Two - Dimensional Electrophoresis: Sample Preparation, Isolation Focalization, Balancing, SDS. Gel staining. DIGE. Differential densitometric analysis of the gel. Mass spectrometry and protein fingerprinting. Maldi, ESI, TOF, Quadrupole, Ion Trap , MS/MS. Analysis in protein identification database (2.5 hours).
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