Efficient enrichment of intact phosphorylated proteins by modified immobilized metal-affinity chromatography

Proteomics. 2005 Dec;5(18):4678-83. doi: 10.1002/pmic.200500002.

Abstract

Phosphoproteome studies are hampered by the lack of methods which allow a comprehensive and fast analysis of intact phosphoproteins. Here we describe an immobilized metal-affinity chromatography (IMAC)-based technique for the enrichment of phosphorylated proteins, which allows recovery of up to 90% of phosphoproteins. This technique is compatible with 2-DE and can be applied to cultured cells and tissues.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Chromatography, Affinity / methods*
  • Electrophoresis, Gel, Two-Dimensional / methods
  • Epithelial Cells / metabolism
  • Humans
  • Mammary Glands, Human / cytology
  • Metals / chemistry
  • Phosphoproteins / isolation & purification*
  • Phosphorylation
  • Proteome / analysis*
  • Smad2 Protein / isolation & purification

Substances

  • Metals
  • Phosphoproteins
  • Proteome
  • SMAD2 protein, human
  • Smad2 Protein