Syllabus
- Introduction. What is proteomics and why ? Proteomics and/or Genomics ? Methods, tools and limitations.
* Basics principles of protein manipulation.
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- Methodology of 2D electrophoresis. Experiment arrangement, sample preparation, isoelectric focusing, PAGE, protein detection and staining. Data validation. 2D electrophoresis and its applications and limitations. Equipment and cost/effectiveness. Technical variations (native electrophoresis, DIGE and fluorescent dyes, blotting).
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- Alternative approaches. Capillary electrophoresis, chromatographic separations, TAP, alternatives to mass spectrometry (antibodies, classical protein sequencing etc). Protein arrays.
- Image analysis of 2D gels. Principles. Reproducibility. Software, Methods, Outputs, Limitations and troubleshooting.
* Preparation of sample for MS analysis. Trypsin and other ways of protein digestion. Peptide extraction from gels and peptide purification.
- Mass spectrometry. MS, MS/MS, MALDI, SELDI, ESI, analyzers, detectors, mass spectra, sensitivity, resolution, peptide identification, gene/protein databases. Special methods (iCATS?.).
Annotation
The lecture is suitable for advanced students of molecular biology and genetics, cellular and developmental biology, parasitology, zoology, as well as other fields. It introduces students to contemporary methods of quantitative and qualitative proteomic analysis using mass spectrometry.
For the year 2023/2024, the lecture has been innovated and expanded to encompass the latest trends in the field, covering the entire range of topics from processing biological samples to data evaluation and the specifics of work in biomedicine. The lecture is delivered by four experienced experts: Jiří Petrák, Petr Man, Karel Harant, and Pavel Talacko.