[Preparation and characterization of monoclonal antibodies against RalB].

Objective: In order to explore the expression of RalB (ras related; GTP binding protein B) in mammal eucaryotic cell, we prepared and characterized monoclonal antibodies against RalB.

Methods: Hybridomas were generated by the fusion with Sp2/0 myelomas and spleen cells, which were from mice immunized with RalB recombinant proteins. The monoclonal antibodies against RalB were then used to identify the expression of RalB in mammal eucaryotic cell, including normal hepatic cell and hepatoma carcinoma cells, by Western blot and Immunohistochemistry.

Results: Two hybridoma cell lines, F001, F002, had been produced, each of which stably secrets antibodies against RalB. Subclass of IgG are both belonged to IgG1. Immunohistochemistry demonstrated that RalB was presented in plasma membrane of hepatoma tissue. Western-blot showed that RalB was expressed in all concerned cell.

Conclusion: The monoclonal antibodies against RalB protein have been successfully prepared, which should provide useful reagent for further investigation into the biological function of RalB.