Native Red Electrophoresis – A new method suitable for separation of native proteins
Tomáš Dráb, Jana Kračmerová, Ivana Tichá, Eva Hanzlíková, Marie Tichá, Helena Ryšlavá, Veronika Hýsková, Pavla Postlerová, Jiřı́ Liberda
Abstract
Tomáš Dráb, Jana Kračmerová, Ivana Tichá, Eva Hanzlíková, Marie Tichá, Helena Ryšlavá, Veronika Hýsková, Pavla Postlerová, Jiřı́ Liberda
Abstract
A new type of native electrophoresis was developed to separate and characterize proteins. In this modification of the native blue electrophoresis, the dye Ponceau Red S is used instead of Coomassie Brilliant Blue to impose uniform negative charge on proteins to enable their electrophoretic separation according to their relative molecular masses. As Ponceau Red S binds less tightly to proteins, in comparison with Coomassie Blue, it can be easily removed after the electrophoretic separation and a further investigation of protein properties is made possible (e.g. an enzyme detection or electroblotting). The tested proteins also kept their native properties (enzyme activity or aggregation state).
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A new type of native electrophoresis was developed to separate and characterize proteins. In this modification of the native blue electrophoresis, the dye Ponceau Red S is used instead of Coomassie Brilliant Blue to impose uniform negative charge on proteins to enable their electrophoretic separation according to their relative molecular masses. As Ponceau Red S binds less tightly to proteins, in comparison with Coomassie Blue, it can be easily removed after the electrophoretic separation and a further investigation of protein properties is made possible (e.g. an enzyme detection or electroblotting). The tested proteins also kept their native properties (enzyme activity or aggregation state).
Key concepts: Coomassie Brilliant Blue, Electrophoresis, Electroblotting, Chromatography, Chemistry, Gel electrophoresis of proteins, Gel electrophoresis, Polyacrylamide gel electrophoresis