Efficient enrichment of intact phosphorylated proteins by modified immobilized metal-affinity chromatography.
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.
Dubrovska
Anna
A
Ludwig Institute for Cancer Research, Biomedical Center, Uppsala University, SE-751 24 Uppsala, Sweden.
Souchelnytskyi
Serhiy
S
D002460:Cell Line; D002846:Chromatography, Affinity / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004847:Epithelial Cells / Q000378:metabolism; D006801:Humans; D042361:Mammary Glands, Human / Q000166:cytology; D008670:Metals / Q000737:chemistry; D010750:Phosphoproteins / Q000302:isolation & purification*; D010766:Phosphorylation; D020543:Proteome / Q000032:analysis*; D051899:Smad2 Protein / Q000302:isolation & purification
D008670:Metals; D010750:Phosphoproteins; D020543:Proteome; C497782:SMAD2 protein, human; D051899:Smad2 Protein