Diethylstilbestrol-hemi-succinate (DES-HS) and diethylstilbestrol-monocarboxylic-propyl-ether (DES-MCPE) were synthesized in different ways. The two kinds of haptens were conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to form the immuno-antigens and the coating-antigen, respectively. The antibody with high titer was obtained after immunising rabbits. The results showed that the antibody had a high affinity to diethylstilbestrol tested by enzyme-linked immunosorbent assay (ELISA) and Western blot. This study lays a basis for further research on the immune test kit for the DES remnant.