Immunoassay for the detection of Carp Vitellogenin.
Immunoassay for the detection of Carp Vitellogenin. This ELISA is developed for a standard test species used in many ecotoxicology laboratories throughout the world, the carp (Cyprinus carpio). The kit may also be used for quantification of Vtg from fathead minnow (Pimephales promelas), another widely used cyprinid test species. Detection of the egg yolk precursor vitellogenin (Vtg) in blood and tissue samples of juvenile and male fish is a simple and sensitive biomarker for endocrine disrupting chemicals (EDCs) with oestrogenic effects. Measurement of Vtg has become an accepted routine screening test for oestrogenic and anti-androgenic effects of EDCs in fish. This Enzyme-Linked Immunosorbent Assay (ELISA) can readily be combined with standard fish toxicology tests developed under the framework of governmental organizations, e.g. the OECD and the US EPA. This ELISA utilizes specific binding between antibodies and vitellogenin (Vtg) to quantify Vtg in samples from carp and fathead minnow. The wells of microplates have been pre-coated with a specific Capture antibody that binds to Vtg in standard and sample added to the wells. A different Vtg-specific Detecting antibody is added to create a sandwich of Vtg and antibody, which is detected with an enzyme-labelled Secondary antibody. The enzyme activity is determined by adding a substrate that gives a colored product, and the color intensity is directly proportional to the amount of Vtg present.