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Complementary Technologies for Phospho-Peptide Enrichment

 

The intrinsically low abundance of critical phosphorylation sites on key proteins requires enrichment techniques for thorough proteomic analysis and understanding of the underlying signaling events within a particular study. Watch this video to learn how two different methods, antibody based immunoaffinty enrichment, and immobilized metal affinity chromatography (IMAC) were employed to offer more complete coverage of the phosphoproteome.

  • PhosphoScan uses PTM specific and motif antibodies to enrich for different classes of phosphopeptides.
  • IMAC methods capture phosphopeptides based on their negative charge and the inherent coordination chemistry dictated by the immobilized metal ion.
  • Each method leads to the identification of thousands of phosphopeptides from drug-treated MKN-45 cells with little overlap between the two different techniques (~ 6%).