Module 1: Introduction |
Concepts: Foundations of Proteomics |
Lab Exercise: Tour of Proteomics Core and Labs |
Skills: Understand Background |
Module 2: Accurate Mass Measurement and Peptide Mass Fingerprinting |
Concepts: Theory of MALDI-TOF-MS, Proteolytic Digestion, Protein Identification using Peptide Masses |
Lab Exercises: Identification of Standard Proteins, Saliva Profiling |
Skills: MALDI Instrument Operation, Protein Denaturation, Tryptic Digestion, Peptide Extraction, Database Searching |
Module 3: Protein Identification using Liquid Chromatography-Tandem Mass Spectrometry Peptide Sequencing |
Concepts: Affinity Purification, Liquid Chromatography, Tandem Mass Spectrometry, In-Gel Digestion, Peptide Sequencing, Database Searching |
Lab Exercise: Analysis of a Protein Complex |
Skills: MALDI MS/MS Instrument Operation, LC-MS/MS Sample Submission, Immunoprecipitation, SDS-PAGE, In-gel Digestion, Database Searching with Tandem Mass Spectra |
Module 4: Proteome Cataloging |
Concepts: Extension of Protein Identification, Proteome Fractionation by Gel-Based or Chromatography-Based Methods, Application to Biomarker Discovery |
Lab Exercise: GeLC-MS/MS and MuDPIT E. coli Proteome Catalogs |
Skills: LC Operation, Fraction Collection, Comparison of Analytical Methods, Quantification of Proteins by Spectral Counting, Summarization of Complex Proteomics Data |
Module 5: Post-Translational Modification Analysis |
Concepts: Molecular Changes by Modification, Separation Techniques for Enrichment, Phosphoproteomic Profiling, Utility in Assessing Drug Response |
Lab Exercise: Analysis of STAT Phosphorylation in K562 Cells |
Skills: Selection of Proteolytic Enzymes Based on Protein Sequence, Affinity Purification of Modified Proteins, Manual Verification of Modification Sites |
Module 6: Protein Quantification |
Concepts: Label-Free Quantification, Relative Quantification using Chemical Labeling (iTRAQ) or Biological Labeling (SILAC), Multiple Reaction Monitoring, Stable Isotope-Labeled Standard Peptides, Biomarker Evaluation |
Lab Exercise: Develop Assays to Quantify Cancer-Related Proteins |
Skills: Assay Development for LC-MRM, Data Analysis |