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.
CITATION STYLE
Schniers, A., Pasing, Y., & Hansen, T. (2019). Lys-C/trypsin tandem-digestion protocol for gel-free proteomic analysis of colon biopsies. In Methods in Molecular Biology (Vol. 1959, pp. 113–122). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9164-8_7
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