A subscription to JoVE is required to view this content. Sign in or start your free trial.
Chapters
- 00:00Introduction
- 00:42Tissue Lysate Preparation
- 01:51Quantification and Quality Check of Tissue Lysates
- 02:33Enzymatic Digestion of Proteins
- 03:57Desalting of Digested Peptides
- 05:16Quantification of Desalted Peptides
- 05:58Labeling of Digested Peptides using iTRAQ Reagents
- 08:14Liquid Chromatography Set-up
- 09:25Mass Spectrometry Set-up
- 12:30Representative Results
- 13:45Conclusions
The described protocol provides an optimized quantitative proteomics analysis of tissue samples using two approaches: label-based and label free quantitation. Label-based approaches have the advantage of more accurate quantitation of proteins, while a label-free approach is more cost-effective and used to analyze hundreds of samples of a cohort.
