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GFP Immunoprecipitation and Sample Preparation for Tandem Mass Tag (TMT) Mass Spectrometry Analysis

By Prosenjit (PPal) Pal, Raja Sekhar Nirujogi, Francesca Tonelli, Dario Alessi, PhD
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Output Details

Description
We describe a method to identify potential interactors of any Green Fluorescent Protein (GFP) tagged protein expressed in mammalian cells by GFP immunoprecipitation coupled to Tandem Mass Tag (TMT) mass spectrometry analysis. As an example, we used a GFP-tagged phosphoRab interactor protein (RILPL1-GFP), and its non-binding mutant (RILPL1 [R293A]-GFP, which cannot interact with phosphorylated Rab proteins) as a control.
Identifier (DOI)
10.17504/protocols.io.eq2ly7kxqlx9/v1
Discovery Team
Team Alessi
Program
Collaborative Research Network
Theme
PD Functional Genomics

Meet the Authors

  • User avatar fallback logo

    Prosenjit (PPal) Pal

    External Collaborator

  • User avatar fallback logo

    Raja Sekhar Nirujogi

    External Collaborator

  • User avatar fallback logo

    Francesca Tonelli

    External Collaborator

  • Dario Alessi, PhD

    Lead PI (Core Leadership): Team Alessi, 

    University of Dundee

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Aligning Science Across Parkinson's
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