Analysis of state-specific phosphorylation of proteins by two-dimensional gel electrophoresis approach |
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Authors: | Kovarova Hana Hajduch Marian Livingstone Mark Dzubak Petr Lefkovits Ivan |
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Affiliation: | Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Rumburska Str 89, CZ-277 21 Libechov, Czech Republic. kovarova@iapg.cas.cz |
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Abstract: | In this paper we focus on the detection of specific state of protein phosphorylation within a complex protein mixture separated by two-dimensional gel electrophoresis followed by immunoblotting. The availability of antibodies that specifically recognize the phosphorylated residue(s) of proteins make this approach feasible as exemplified by the study of the regulatory mechanisms of the cell cycle. The major advantage of the presented approach is its relative simplicity and sensitivity that allows specific detection of protein phosphorylation and distinguishes different phosphorylation sites of target protein. Current findings demonstrate that this method represents a reasonable alternative to the use of other tools to study protein phosphorylation. |
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Keywords: | Protein phosphorylation Roscovitine |
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