2. Academic Validation
  3. Protocol for identification of protein citrullination by immunoprecipitation followed by mass spectrometry

Protocol for identification of protein citrullination by immunoprecipitation followed by mass spectrometry

  • STAR Protoc. 2026 Jan 12;7(1):104326. doi: 10.1016/j.xpro.2025.104326.
Alexandra Indeglia 1,  Hsin-Yao Tang 2
Affiliations

Affiliations

  • 1 Program 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. Electronic address: [email protected].
  • 2 Program in Molecular and Cellular Oncogenesis, The Wistar Institute, Philadelphia, PA 19104, USA. Electronic address: [email protected].
PMID: 41528850 DOI: 10.1016/j.xpro.2025.104326
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.

Keywords

Genomics; Molecular biology; Protein biochemistry; Proteomics.

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