Production of rat monoclonal antibodies against RNA-binding protein Hzf.

The hematopoietic zinc finger protein, Hzf, is induced in response to DNA damage or by Arf tumor suppressor in a p53-dependent manner. Recent studies have revealed that Hzf is an RNA-binding protein that regulates localization and translation of specific mRNA. The RNA-binding activity of Hzf is required for the functions of cerebellar purkinje cells and adipocytes, although their molecular mechanisms underlying the mRNA regulation largely remain unknown. To further investigate the molecular function of Hzf, we raised two rat monoclonal antibodies (MAb) against a peptide corresponding to the C-terminal region of the mouse/human Hzf protein. Both MAbs reacted with the native protein expressed in mammalian cells, and were highly efficient in detecting endogenous Hzf by immunoblotting, immunoprecipitation, and immunofluorescence. These MAbs should therefore be useful for further analysis of molecular functions of the Hzf protein and for identification of Hzf-binding proteins.