PUBLICATION
Development of an ELISA for vitellogenin in whole body homogenate of zebrafish (Danio rerio)
- Authors
- Holbech, H., Andersen, L., Petersen, G.I., Korsgaard, B., Pedersen, K.L., and Bjerregaard, P.
- ID
- ZDB-PUB-010912-6
- Date
- 2001
- Source
- Comparative biochemistry and physiology. Toxicology & pharmacology : CBP 130(1): 119-131 (Journal)
- Registered Authors
- Andersen, Lene, Bjerregaard, Poul, Holbech, Henrik, Korsgaard, Bodil, Pedersen, Knud L., Petersen, Gitte I.
- Keywords
- biomarker; ELISA; endocrine disrupters; ethinylestradiol; lipovitellin; vitellogenin; western blotting; zebrafish
- MeSH Terms
-
- Animals
- Antibodies
- Blotting, Western
- Chromatography, Gel
- Chromatography, Ion Exchange
- Egg Proteins
- Egg Proteins, Dietary/immunology
- Egg Proteins, Dietary/isolation & purification
- Electrophoresis
- Enzyme-Linked Immunosorbent Assay/methods*
- Ethinyl Estradiol/pharmacology
- Male
- Reproducibility of Results
- Vitellogenins/analysis*
- Vitellogenins/drug effects
- Vitellogenins/immunology
- Vitellogenins/isolation & purification
- Zebrafish/metabolism*
- PubMed
- 11544148 Full text @ Comp. Biochem. Physiol. C Toxicol. Pharmacol.
Citation
Holbech, H., Andersen, L., Petersen, G.I., Korsgaard, B., Pedersen, K.L., and Bjerregaard, P. (2001) Development of an ELISA for vitellogenin in whole body homogenate of zebrafish (Danio rerio). Comparative biochemistry and physiology. Toxicology & pharmacology : CBP. 130(1):119-131.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping