title: Ink-native electrophoresis: an alternative to blue-native electrophoresis more suitable for in-gel detection of enzymatic activity.
abstract: Blue-native electrophoresis (BNE) is a useful technique for analyzing protein complexes, but the Coomassie brilliant blue (CBB) dye used in BNE often hampers in-gel detection of enzymatic activity. Here we report an improved method, termed ink-native electrophoresis (INE), in which Pelikan 4001 fountain pen ink is used as a charge-shifting agent instead of CBB. INE is more suitable than BNE for in-gel detection of protein kinase activity after polyacrylamide gel electrophoresis (PAGE), and its performance in protein complex separation is comparable to that of conventional BNE. INE may provide a powerful tool to isolate and analyze various protein complexes.
authors.affiliation: Department of Life Sciences, Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa 761-0795, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Kaneko
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Sueyoshi
authors.forename: Isamu
authors.initials: I
authors.lastname: Kameshita
authors.forename: Atsuhiko
authors.initials: A
authors.lastname: Ishida
mesh_terms: D004586:Electrophoresis / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods*; D005782:Gels; D007281:Ink; D011494:Protein Kinases / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D005782:Gels; D011494:Protein Kinases
keywords: Autophosphorylation; Calmodulin; In-gel assay; Native PAGE; Protein kinase