Purification and establishment of ELISA for vitellogenin of Japanese sardine (Sardinops melanostictus) |
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| Institution: | 1. Environmental and Fisheries Sciences Division, Northwest Fisheries Science Center, National Marine Fisheries Service, National Oceanic and Atmospheric Administration, 2725 Montlake Blvd East, Seattle, WA 98112, USA;2. School of Aquatic and Fishery Sciences, University of Washington, 1122 NE Boat St, Seattle, WA 98105, USA;3. Center for Reproductive Biology, Washington State University, PO Box 647521, Pullman, WA 99164, USA;4. Ocean Associates Inc., Under Contract to Northwest Fisheries Science Center, National Marine Fisheries Service, National Oceanic and Atmospheric Administration, 2725 Montlake Blvd East, Seattle, WA 98112, USA;1. Deakin Genomics Centre, Deakin University, Geelong 3220, Victoria, Australia;2. School of Life and Environmental Sciences, Deakin University, Geelong 3220, Victoria, Australia;3. School of Science, Monash University Malaysia, Bandar Sunway 47500, Selangor, Malaysia |
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| Abstract: | Two female-specific serum proteins (FSSPs) were detected immunologically in estradiol-treated Japanese sardine Sardinops melanostictus. The major FSSP was demonstrated to be a high molecular estradiol-inducible glycolipophosphoprotein with an immunological relation to a major yolk protein, and was suggested to be vitellogenin (VTG). VTG was purified using negative immunoaffinity chromatography. The isolated VTG was used for raising the specific antiserum against VTG. A homologous enzyme-linked immunosorbent assay (ELISA) was developed using the antiserum and the isolated VTG. The sensitivity range of the ELISA was 44 ng/ml to 2670 ng/ml of VTG concentration for the conditions used in our investigation. |
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