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Efficient enrichment of intact phosphorylated proteins by modified immobilized metal‐affinity chromatography
Author(s) -
Dubrovska Anna,
Souchelnytskyi Serhiy
Publication year - 2005
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200500002
Subject(s) - affinity chromatography , phosphorylation , chemistry , chromatography , proteomics , protein phosphorylation , biochemistry , enzyme , protein kinase a , gene
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.