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Preparation and Application of a Decellularized Extracellular Matrix for Identification of ADAMTS Substrates

  • Rahel SchnellmannEmail author
  • Ruth Chiquet-Ehrismann
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2043)

Abstract

Here we describe the use of a decellularized ECM produced in vitro by BALB/c 3T3 fibroblasts for the identification of ADAMTS substrates. Seeding of ADAMTS protease-producing HEK cells on top of the cell-free ECM followed by analysis of the conditioned medium by liquid chromatography tandem mass spectrometry (LC-MS/MS), allows for screening of ADAMTS substrates without prior purification of full-length protease.

Key words

Decellularized extracellular matrix Substrate identification ADAMTS protease Mass spectrometry 

Notes

Acknowledgments

This work was supported by a grant from The Swiss National Science Foundation (SNF, Project funding in biology and medicine (division III)) grant no. 31003A_156740.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Friedrich Miescher Institute for Biomedical ResearchBaselSwitzerland
  2. 2.Faculty of ScienceUniversity of BaselBaselSwitzerland
  3. 3.Department of Chemical and Biomolecular Engineering, The Institute for NanoBioTechnologyJohns Hopkins UniversityBaltimoreUSA

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