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Updated: Jul 23, 2026

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation

Published on: September 24, 2012

Protocolo para la identificación de la citrulinación de proteínas mediante inmunoprecipitación seguida de

Alexandra Indeglia¹, Hsin-Yao Tang²

¹Program in Molecular and Cellular Oncogenesis, The Wistar Institute, Philadelphia, PA 19104, USA; Graduate Group in Biochemistry and Molecular Biophysics, the University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.

|January 13, 2026

Abstract:

Protein citrullination occurs through the deimination of peptidyl-arginine to yield peptidyl-citrulline by one of the peptidyl-arginine deiminase (PAD) family members. This protocol identifies citrullinated protein residues using immunoprecipitation followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). We describe steps for identification of citrullination modifications in vitro or in cell culture, immunoprecipitation of nuclear citrullinated proteins, and identification of citrullinated residues by mass spectrometry (MS). This protocol applies to both recombinant protein assays and in vitro cell culture assays. For complete details on the use and execution of this protocol, please refer to Indeglia et al.¹.

Palabras clave:

Genómica Biología molecular Bioquímica de proteínas Proteómica

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