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Proteomics for Biomarker Discovery pp 113–122Cite as

Lys-C/Trypsin Tandem-Digestion Protocol for Gel-Free Proteomic Analysis of Colon Biopsies

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1959))

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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Acknowledgments

We thank Ilona Urbarova and Jack-Ansgar Bruun for fruitful discussions, as well as Prof Jon Florholmen and Rasmus Goll for supplying colon biopsies. Our work was supported by a grant from the North Norway Regional Health Authorities.

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Authors and Affiliations

  1. Natural Products and Medicinal Chemistry Research Group, Department of Pharmacy, UiT—The Arctic University of Norway, Tromsø, Norway

    Armin Schniers & Terkel Hansen

  2. Tromsø Endocrine Research Group, Department of Clinical Medicine, UiT—The Arctic University of Norway, Tromsø, Norway

    Yvonne Pasing

  3. Division of Internal Medicine, University Hospital of North Norway, Tromsø, Norway

    Yvonne Pasing

Authors
  1. Armin Schniers
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  2. Yvonne Pasing
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  3. Terkel Hansen
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Corresponding author

Correspondence to Terkel Hansen .

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Editors and Affiliations

  1. Université Grenoble Alpes, CEA, Inserm, BGE U1038, Grenoble, France

    Virginie Brun

  2. Université Grenoble Alpes, CEA, Inserm, BGE U1038, Grenoble, France

    Yohann Couté

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Schniers, A., Pasing, Y., Hansen, T. (2019). Lys-C/Trypsin Tandem-Digestion Protocol for Gel-Free Proteomic Analysis of Colon Biopsies. In: Brun, V., Couté, Y. (eds) Proteomics for Biomarker Discovery. Methods in Molecular Biology, vol 1959. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9164-8_7

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  • DOI: https://doi.org/10.1007/978-1-4939-9164-8_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9163-1

  • Online ISBN: 978-1-4939-9164-8

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