STAR Protocols (Dec 2020)

Organoid Sample Preparation and Extraction for LC-MS Peptidomics

  • Emily L. Miedzybrodzka,
  • Rachel E. Foreman,
  • Sam G. Galvin,
  • Pierre Larraufie,
  • Amy L. George,
  • Deborah A. Goldspink,
  • Frank Reimann,
  • Fiona M. Gribble,
  • Richard G. Kay

Journal volume & issue
Vol. 1, no. 3
p. 100164

Abstract

Read online

Summary: This protocol describes the peptidomic analysis of organoid lysates, FACS-purified cell populations, and 2D culture secretions by liquid chromatography mass spectrometry (LC-MS). Currently, most peptides are quantified by ELISA, limiting the peptides that can be studied. However, an LC-MS-based approach allows more peptides to be monitored. Our group has previously used LC-MS for tissue peptidomics and secretion of enteroendocrine peptides from primary culture. Now, we extend the use to organoid models.For complete details on the use and execution of this protocol, please refer to Goldspink et al. (2020).

Keywords