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Using Co-immunoprecipitation and Shotgun Mass Spectrometry for Protein-Protein Interaction Identification in Cultured Human Oligodendrocytes

  • Bradley Smith
  • Daniel Martins-de-Souza
  • Mariana Fioramonte
Protocol
Part of the Neuromethods book series (NM, volume 144)

Abstract

Protein complexes are responsible for major biological processes in cells. Identifying a protein’s interaction partners reveals many different aspects related to the target such as its function, regulation, and mechanisms of action. Co-immunoprecipitation (co-IP) followed by shotgun proteomics is a powerful technique for capturing and identifying endogenous protein complexes. Although it is a simple concept, performing the experiment can be challenging due to the number of steps and the diversity of available protocols. Here we present a certified and detailed protocol to perform co-IP experiments in cultured human oligodendrocytes cells.

Key words

Co-immunoprecipitation Mass spectrometry Protein-protein interaction Interactome Proteomics 

Notes

Acknowledgments

Research was funded by FAPESP (São Paulo Research Foundation, grants 2016/18715-4 and 2016/07948-8).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Bradley Smith
    • 1
  • Daniel Martins-de-Souza
    • 1
    • 2
  • Mariana Fioramonte
    • 1
  1. 1.Laboratory of Neuroproteomics, Department of Biochemistry and Tissue Biology, Institute of BiologyUniversity of Campinas (UNICAMP)CampinasBrazil
  2. 2.Instituto Nacional de Biomarcadores em Neuropsiquiatria (INBION), Conselho Nacional de Desenvolvimento Cientifico e TecnologicoSao PauloBrasil

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