Crucian carp () vitellogenin (Vtg) is a commonly used biomarker for the detection of estrogenic activity in aquatic environments. In this study, gel filtration combined with ion-exchange chromatography and selective precipitation combined with ion-exchange chromatography were used to purify lipovitellin (Lv) from ovarian homogenates of crucian carp. Lv was identified as a phospholipogly coprotein with an apparent molecular weight of~521 kD and separated into two major polypeptides corresponding to~117 and~103 kD by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The results of the western blot showed that both anti-goldfish () Lv antibodies had good cross-reaction with crucian carp Lv. Using anti-goldfish Lv antibody, anti-zebrafish Lv antibody, and purified crucian carp Lv, two sandwich enzyme-linked immunosorbent assays (ELISAs) were established. The results showed that the binding curves of goldfish Lv and crucian carp Lv basically overlapped. The sandwich ELISA developed using anti-goldfish Lv antibodies and purified crucian carp Lv had a working range form 15.6 to 1000 ng/mL and a detection limit of 6.8 ng/mL, which was significantly lower than that of the anti-zebrafish Lv antibody-based ELISA and other ELISAs previously reported for crucian carp Vtg. Therefore, we suggest that it is a reliable tool for the quantification of crucian carp Vtg.