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Updated: Aug 28, 2025

Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics
Published on: July 6, 2014
Endogenous protein interactomes resolved through immunoprecipitation-coupled quantitative proteomics in cell lines
Raman Kumar1, Karthik S Kamath2, Luke Carroll2
1Adelaide Medical School and the Robinson Research Institute, University of Adelaide, Adelaide, SA 5005, Australia.
Abstract:
Immunoprecipitation (IP) of endogenously expressed proteins is one of the most biologically relevant techniques to identify protein-protein interactions. We describe an adaptable IP protocol reliant on a specific antibody to the target protein. We detail a quantitative proteomics workflow for the unbiased identification of co-immunoprecipitating proteins, known collectively as an interactome. This includes protocols for the tryptic digestion, Tandem Mass Tag labeling and fractionation of peptides, and their identification and quantification using liquid chromatography-mass spectrometry including computational and statistical analysis. For complete details on the use and execution of this protocol, please refer to Johnson et al. (2020).
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