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

Phosphoproteome studies are hampered by the lack of methods which allow a comprehensive Received: January 4, 2005 and fast analysis of intact phosphoproteins. Here we describe an immobilized metal-affinity Revised: April 16, 2005 chromatography (IMAC)-based technique for the enrichment of phosphorylated proteins, which Accepted: April 25, 2005 allows recovery of up to 90% of phosphoproteins. This technique is compatible with 2-DE and can be applied to cultured cells and tissues.