A new ELISA for the three-spined stickleback (Gasterosteus aculeatus L.) spiggin, using antibodies against synthetic peptide - Ineris - Institut national de l'environnement industriel et des risques Access content directly
Journal Articles Comparative Biochemistry and Physiology - Part C: Toxicology and Pharmacology Year : 2008

A new ELISA for the three-spined stickleback (Gasterosteus aculeatus L.) spiggin, using antibodies against synthetic peptide

Abstract

The aim of this study was to develop an enzyme-linked immunosorbent (ELISA) assay to quantify spiggin in the three-spined stickleback. Spiggin is a glue protein produced in the kidney of male three-spined stickleback under the control of androgens during the breeding period. Disturbances of spiggin production in male fish and abnormal induction of spiggin in female fish are considered as valuable biomarkers of exposure to (anti-)androgenic chemicals. Polyclonal antibodies against a peptide sequence of spiggin (HRD-16) were used and the specificity of the antibodies was verified by Western blotting and direct ELISA experiments. By using HRD-16 antibodies and spiggin standard preparation, a competitive ELISA was set-up and validated. This assay appears sensitive, with a detection limit of 0.5 U/mL, and specific, as shown by the competition curves, obtained by serial dilution of male and female kidney homogenates, that were parallel to the spiggin standard curves. The ability of the spiggin ELISA to quantify spiggin induction was achieved by exposing male and female three-spined sticklebacks to 0.1 and 1 micro g/L of methyltestosterone. The results show a significant dose-dependent induction of spiggin in methyltestosterone-exposed female fish compared to controls.
Fichier principal
Vignette du fichier
2007-217_post-print.pdf (235.86 Ko) Télécharger le fichier
Origin : Files produced by the author(s)
Loading...

Dates and versions

ineris-00961919 , version 1 (20-03-2014)

Identifiers

Cite

Wilfried Sanchez, Carolyne Goin, François Brion, P.E. Olsson, Anders Goksoyr, et al.. A new ELISA for the three-spined stickleback (Gasterosteus aculeatus L.) spiggin, using antibodies against synthetic peptide. Comparative Biochemistry and Physiology - Part C: Toxicology and Pharmacology, 2008, 147 (1), pp.129-137. ⟨10.1016/j.cbpc.2007.08.007⟩. ⟨ineris-00961919⟩

Collections

INERIS
230 View
415 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More