Gilthead Seabream (Sparus aurata) Vitellogenin: Purification, Partial Characterization, and Validation of an Enzyme-Linked Immunosorbent Assay (ELISA)
| Autor: | Massimo Nabissi, Oliana Carnevali, A.M. Polzonetti-Magni, R. Carletta, Gilberto Mosconi |
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| Rok vydání: | 1998 |
| Předmět: |
Male
chemistry.chemical_classification Estradiol Molecular mass Blotting Western Ion chromatography Reproducibility of Results Enzyme-Linked Immunosorbent Assay Biology Perciformes Blot Vitellogenins Vitellogenin Endocrinology Enzyme Liver Biochemistry chemistry Concanavalin A Sephadex biology.protein Animals Electrophoresis Polyacrylamide Gel Animal Science and Zoology Polyacrylamide gel electrophoresis |
| Zdroj: | General and Comparative Endocrinology. 110:252-261 |
| ISSN: | 0016-6480 |
| Popis: | VTG was purified from seabream Sparus aurata plasma by ion exchange chromatography on a DEAE-Sepharose column. The vitellogenin was characterized and its properties were determined. The molecular mass of the native form, obtained by Sephadex G-200 column, was around 450 kDa, whereas an apparent molecular mass of 180 kDa was detected by electrophoresis under denaturing and reducing conditions, suggesting a dimeric form for the native protein. The presence of carbohydrates was determined using concanavalin A, while the presence of phosphate groups was detected by Stains-all, a cationic stain. These data together with the sex specificity, the estrogen inducibility, and the cross-reactivity of the abVTG against the major yolk proteins identifies this protein as vitellogenin. The validated ELISA was used for a rapid and reliable measurement of plasma VTG changes related with those of estradiol-17beta in female broodstock. |
| Databáze: | OpenAIRE |
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