Molecular analytical methods - PROTEOMICA (2013/2014)

Course code
4S02774
Credits
4
Academic sector
CHIM/01 - ANALYTICAL CHEMISTRY
Language of instruction
Italian
Teaching is organised as follows:
Activity Credits Period Academic staff Timetable
teoria 3 II semestre Daniela Cecconi
laboratorio 1 II semestre Daniela Cecconi

Lesson timetable

II semestre
Activity Day Time Type Place Note
teoria Tuesday 8:30 AM - 10:30 AM lesson Lecture Hall H  
teoria Thursday 11:30 AM - 1:30 PM lesson Lecture Hall H  

Learning outcomes

Proteomics is the large-scale study of proteins. It is an emerging scientific research which can be applied to food (to find allergen for example) and plants research (for example to study plant pathogen response).

Syllabus

PROTEOMICS

Analytical proteomics (LM9+LM7):

1. Introduction.

2. Sample preparation.

3. Gel-based separation.

4. 2D gel Image analysis.

5. Gel-free separation.

6. Pre-fractionation of low abundance proteins.

7. Analysis of proteins/peptides by Mass Spectrometry.

8. Analysis of proteins/peptides by Tandem Mass Spectrometry.

9. Analysis of mass spectra and database search for protein identification.

10. Differential and Quantitative proteomics.

11. Characterization of post-translational modifications.

12. Proteomics in agro-food and biomedical areas.



Laboratory (1 CFU)

Day 1:
Rehydration of 7 cm IPG with protein extract
Preparation of buffers and solutions for 2-DE analysis
Polymerization of 12% T gel
Isoelectrofocusing by using PROTEAN® IEF Cell (Bio-Rad)


Day 2:
IPG strip equilibration
Interfacing with the second dimension
Second dimension by Mini-PROTEAN® 3 Dodeca™ Cell (Bio-Rad)
Staining of 2-DE maps with Coomassie and/or SyproRuby

Day 3:
Destaining and acquisition of images by VersaDoc™ MP 1000 system (Bio-Rad)
Image analysis by PDQuest software (Bio-Rad) (classroom gamma)
Analysis of mass spectra and database searching with Mascot

Assessment methods and criteria

written

Mandatory attendance to laboratory.