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

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

Team

Team Alessi

Program

Collaborative Research Network

Theme

PD Functional Genomics

Meet the Authors

Prosenjit Pal, PhD

Key Personnel: Team Alessi,

University of Dundee
Raja Sekhar Nirujogi, PhD

Key Personnel: Team Alessi,

University of Dundee
Francesca Tonelli, PhD

Key Personnel: Team Alessi,

University of Dundee
Dario Alessi, PhD

Lead PI (Core Leadership): Team Alessi,

University of Dundee

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

Output Details

Meet the Authors

Prosenjit Pal, PhD

Raja Sekhar Nirujogi, PhD

Francesca Tonelli, PhD

Dario Alessi, PhD

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