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Proteomic Profile of EPS-Urine through FASP Digestion and Data-Independent Analysis
This content is Free Access.
1Research Centre for Advanced Biochemistry and Molecular Biology, Department of Experimental and Clinical Medicine,Magna Graecia University of Catanzaro, 2Department of Health Science,Magna Graecia University of Catanzaro, 3Romolo Hospital, 4Department of Experimental and Clinical Medicine,Magna Graecia University of Catanzaro
Chapters
- 00:00Introduction
- 00:55Protocol
- 01:25Protocol: FASP Digestion
- 05:29Protocol: SCX Purification
- 08:08Protocol: Protein Quantifcation by External Standard
- 09:23Protocol: NanoLC-MS/MS and DIA Analysis
- 11:31Protocol: High-pH C18 Fractionation for Spectral Library
- 13:09Protocol: Data Analysis
- 13:35Expected Results
- 13:53Conclusion
Here, we present an optimized on-filter digestion protocol with detailed information about the following: protein digestion, peptide purification and data independent acquisition analysis. This strategy is applied to the analysis of expressed prostatic secretions-urine samples and allows high proteome coverage and low missing value label-free profiling of the urinary proteome.
