ObjectiveTo generate monoclonal antibodies against novel zinc finger protein ZBTB45 of mouse for in vitro and in vivo immunoassay of the protein, so as to contribute to the study on its biological functions. MethodsMulti-epitope antigen of ZBTB45 was prepared by prokaryotic fusion expression and was used to immunize BALB/c mice, and then the splenocytes of mice were fused with the myeloma cells. The resultant fused cells were subjected to screening culture, ELISA assay and subcloning by limited dilution to establish monoclonal hybridomas secreting anti-ZBTB45 antibody. The antibody was purified by affinity chromatography with protein A column from the ascites of hybridoma-bearing mice, and was tested by Western blotting analysis, immunoprecipitation, and immunohistochemistry. ResultsWe successfully established 6 cell lines of monoclonal hybridoma secreting anti-ZBTB45 antibody, of which monoclonal antibodies 3E5, 2G4, 4D2, and 1D4 were applicable for Western blotting analysis and immunoprecipitation, and 3E5, 6D8, and 8E3 were applicable for detecting endogenous ZBTB45 by immunohistochemistry, with a satisfying sensitivity and specificity.ConclusionWe have for the first time successfully generated monoclonal antibodies against mouse ZBTB45, which are applicable in various immunoassays for ZBTB45 both in vitro and in vivo.