Abstract
The yolk protein, lipovitellin (Lv) was purified from ovaries of mature female zebrafish ( Danio rerio) by gel filtration and anion exchange chromatography. Polyclonal antibodies against Lv were raised in rabbits. Anti-Lv IgG was purified by affinity chromatography. SDS-PAGE followed by Western blotting was performed to analyse the specificity of the antibody and the immunological similarities between Lv and vitellogenin (Vtg). Anti-Lv IgG was used to develop a direct non-competitive sandwich ELISA to measure Vtg concentrations of whole body homogenate (WBH) in zebrafish. The intra- and interassay variabilities were 5.8% and 10.4%, respectively. The sensitivity was 0.2 ng Vtg ml −1 and the practical detection limit was 40 ng Vtg g −1 fish (wet weight). Adult male zebrafish were exposed to a nominal water concentration of 10 ng l −1 of ethinylestradiol (EE 2) in a semi-static exposure system for 7 days. Compared with the control group, exposure to 10 ng EE 2 l −1 induced a 200-fold increase in Vtg levels.
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