Production of mouse recombinant JAK2 protein

Abstract

This work describes the production of JAK2 by recombinan, methods, using a baculovirus expression vector. Sf9 cells from the ovary tissue of Spodoptera frugiperda were infected with an optimal virus dilution of 10plIml medium and harvest was done at 72 h postinfection. Approximately 5 x 107cells were infected, extracts were collected, ultrafiltered and subjected to ionic exchange chromatography on DEAE-Sephacel, and JAK2 was partially purified by a continuos ionic gradient (1 0-500 mM NaCI). JAK2 was identified by Western blot with a polyclonal antibody raised in rabbits against 758-776 residues from murine JAK2. Results showed that the protein obtained by this method is in phosphorylatedl activated state.