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Identifying Myc interactors

Romina Ponzielli1, William B Tu, Igor Jurisica

  • 1Ontario Cancer Institute, The Campbell Family Institute for Cancer Research, Toronto, ON, Canada.

Methods in Molecular Biology (Clifton, N.J.)
|September 6, 2013
PubMed

Insights

This chapter details co-immunoprecipitation (Co-IP) and in vitro pull-down assays for identifying Myc protein interactions. These methods help validate potential Myc interactors found through in silico screening.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Cell Biology

Background:

  • Myc proteins are crucial transcription factors involved in cell growth and proliferation.
  • Understanding Myc's protein-protein interactions is key to elucidating its regulatory functions.
  • Existing methods for identifying protein interactions require detailed protocols for validation.

Purpose of the Study:

  • To provide detailed methodologies for co-immunoprecipitation (Co-IP) and in vitro pull-down assays.
  • To enable researchers to investigate Myc protein interactions within cellular contexts.
  • To offer strategies for screening and validating Myc-interacting proteins.

Main Methods:

  • Co-immunoprecipitation (Co-IP) from various cellular fractions (whole-cell, cytoplasmic, nuclear).
  • In vitro pull-down assays to assess direct and indirect protein-protein interactions.
  • In silico strategies for prioritizing potential Myc interactors.

Main Results:

  • Detailed protocols for performing Co-IP and in vitro pull-down assays are presented.
  • Methods allow for the identification of Myc interacting partners.
  • The assays can distinguish between direct and indirect protein interactions.

Conclusions:

  • Co-IP and in vitro pull-down assays are essential for studying Myc protein interactions.
  • These techniques, combined with in silico screening, provide a robust approach for Myc interactor discovery and validation.

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