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

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.