Details zur Publikation |
| Kategorie | Textpublikation |
| Referenztyp | Zeitschriften |
| DOI | 10.1016/S1532-0456(01)00194-6 |
| Titel (primär) | Development and validation of a Homologous Zebrafish (Danio rerio HAMILTON-BUCHANAN) vitellogenin Enzyme-Linked Immunosorbent Assay (ELISA) and its application for studies on estrogenic chemicals |
| Autor | Fenske, M.; van Aerle, R.; Brack, S.; Tyler, C.R.; Segner, H. |
| Journal / Serie | Comparative Biochemistry and Physiology C-Toxicology & Pharmacology |
| Erscheinungsjahr | 2001 |
| Department | OEC; COE |
| Band/Volume | 129 |
| Seite von | 217 |
| Seite bis | 232 |
| Sprache | englisch |
| Keywords | Enzyme-linked immunosorbent assay (ELISA); Environmental estrogens; 17α-Ethinylestradiol (EE2); Homologous antibody; Vitellogenin (VTG); Zebrafish |
| Abstract | Vitellogenin (VTG) was isolated by anion exchange chromatography from plasma of female zebrafish (Danio rerio) induced with 17α-ethinylestradiol (EE2). The purity of the VTG isolate was confirmed by polyacrylamide gel electrophoresis (SDS-PAGE). Purified VTG was used to raise polyclonal antibodies in rabbits and the specificity of the antisera for VTG confirmed by Western blot analysis of plasma proteins separated by SDS-PAGE. The antibodies cross-reacted with two proteins in the plasma of female zebrafish, with molecular masses of approximately 142 and 171 kDa. No cross-reactivity was observed with any other plasma proteins. A competitive enzyme-linked immunosorbent assay (ELISA) was developed using the polyclonal zebrafish VTG (z-VTG) antibodies and purified z-VTG as ligand and standard, respectively. The z-VTG ELISA was sensitive with a detection limit of between 2.0 and 3.0 ng purified VTG/ml, and a working range between 3 and 500 ng/ml (30–85% binding). The ELISA demonstrated precision, with inter- and intra-assay variations of 7.5±2.7 and 4.9±1.4%, respectively. Plasma from adult zebrafish and whole body homogenates from juvenile zebrafish diluted parallel with the z-VTG standard in the ELISA, validating the assay for quantifying z-VTG in both of these tissues. Exposure of adult male zebrafish to EE2 via water induced a concentration-dependent induction of VTG with a lowest observed effect concentration (LOEC) ≤1.67 ng EE2/l (for a 21-day exposure). The homologous z-VTG ELISA provides a valuable tool for the study of environmental estrogens in zebrafish. |
| dauerhafte UFZ-Verlinkung | https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=6311 |
| Fenske, M., van Aerle, R., Brack, S., Tyler, C.R., Segner, H. (2001): Development and validation of a Homologous Zebrafish (Danio rerio HAMILTON-BUCHANAN) vitellogenin Enzyme-Linked Immunosorbent Assay (ELISA) and its application for studies on estrogenic chemicals Comp. Biochem. Physiol. C-Toxicol. Pharmacol. 129 , 217 - 232 |
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