Organoid Sample Preparation and Extraction for LC-MS Peptidomics

STAR Protoc. 2020 Nov 25;1(3):100164. doi: 10.1016/j.xpro.2020.100164. eCollection 2020 Dec 18.

Abstract

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: Mass Spectrometry; Organoids; Proteomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Chromatography, Liquid
  • Flow Cytometry
  • Humans
  • Organoids / metabolism*
  • Peptides / chemistry
  • Peptides / isolation & purification
  • Peptides / metabolism*
  • Proteomics / methods*
  • Tandem Mass Spectrometry*

Substances

  • Peptides