In the base of truncated Flavanone-3-hydroxylase gene (TrF3H) achieved from full-length cDNA library of tartary buckwheat which was in seed-filling period,prokaryotic expression vector pET47b-TrF3H with truncated coding region of F3H（TrF3H）was constructed and transformed into E.coli Rosetta (DE3) plysS.After the overexpressed target protein purified by cobalt chelating chromatography,the high titer polyclonal antiserum raised against rabbit was obtained.The results demonstrated that the TrF3H had been expressed in E.coli in the form of inclusion bodies and western blotting analysis showed the raised antibody could specifically react with the antigen while native F3H existed in immature seed protein.These results laid the basis for further exploration of the functions of F3H in tartary buckwheat.