Unit BIOCHEMICAL METHODS
- Course
- Biology
- Study-unit Code
- GP004082
- Curriculum
- Bionutrizionistico
- 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
- Academic discipline
- 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).