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Lys-C/Trypsin Tandem-Digestion Protocol for Gel-Free Proteomic Analysis of Colon Biopsies
Armin Schniers1, Yvonne Pasing2,3, Terkel Hansen4
1Natural Products and Medicinal Chemistry Research Group, Department of Pharmacy, UiT-The Arctic University of Norway, Tromsø, Norway.
Abstract:
The protocol presented was specifically optimized for in-depth analysis of the human colon mucosa proteome. After cell lysis in a sodium deoxycholate/urea buffer, a tandem digestion with Lys-C and trypsin was performed. Prior to LC-MS/MS analysis, peptides were TMT-labeled and fractionated by high pH reversed-phase spin columns. This protocol is a powerful, reproducible, sample-saving, and cost-effective option when an in-depth quantitative proteome analysis is desired.
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