Plasma Vitellogenin Levels during the Annual Reproductive Cycle of the Female Rainbow Trout ( Oncorhynchus mykiss ): Establishment and Validation of an ELISA
Identifieur interne : 001949 ( Main/Merge ); précédent : 001948; suivant : 001950Plasma Vitellogenin Levels during the Annual Reproductive Cycle of the Female Rainbow Trout ( Oncorhynchus mykiss ): Establishment and Validation of an ELISA
Auteurs : E. Bon [France] ; U. Barbe [France] ; J. Nu Ez Rodriguez [Côte d'Ivoire] ; B. Cuisset [France] ; C. Pelissero [France] ; J. P Sumpter [Royaume-Uni] ; Françoise Le Menn [France]Source :
- Comparative Biochemistry and Physiology, Part B: Biochemistry and Molecular Biology [ 1096-4959 ] ; 1997.
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Abstract
Rainbow trout, Oncorhynchus mykiss, vitellogenin (Vtg) was purified from plasma of E2-treated male by direct anion exchange chromatography and some of its biochemical characteristics were studied. Our results demonstrated that, under SDS-PAGE conditions, rainbow trout Vtg was composed of two molecular forms of 390 and 176 kDa representing, respectively, the dimeric form and the monomeric form of the molecule. The purified Vtg was used to raise a polyclonal antibody for Vtg (anti-Vtg). Using this anti-Vtg, a competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantification of rainbow trout Vtg. The practical sensitivity range of this ELISA was 20–320 ng/ml (80–20% of binding) and the detection limit was 9 ng/ml. The intra- and the inter-assay coefficients of variation (at 50% of binding) were estimated at 1.8% (n = 10) and 3.9% (n = 13), respectively. This ELISA was validated by detecting changes in Vtg levels in rainbow trout at different physiological stages, as well as in 2-year-old female rainbow trout throughout the reproductive cycle.
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DOI: 10.1016/S0305-0491(96)00252-0
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P" last="Sumpter">J. P Sumpter</name></author><author><name sortKey="Le Menn, F" sort="Le Menn, F" uniqKey="Le Menn F" first="F" last="Le Menn">Françoise Le Menn</name><affiliation><country>France</country><placeName><settlement type="city">Bordeaux</settlement><region type="region" nuts="2">Nouvelle-Aquitaine</region><region type="old region" nuts="2">Aquitaine</region></placeName><orgName type="university" n="3">Université Bordeaux I</orgName><orgName type="institution" wicri:auto="newGroup">Université de Bordeaux</orgName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:941BA0C804C416A015013F99C10A95DE34D15685</idno><date when="1997" year="1997">1997</date><idno type="doi">10.1016/S0305-0491(96)00252-0</idno><idno type="url">https://api.istex.fr/document/941BA0C804C416A015013F99C10A95DE34D15685/fulltext/pdf</idno><idno type="wicri:Area/Istex/Corpus">000C89</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000C89</idno><idno type="wicri:Area/Istex/Curation">000C87</idno><idno type="wicri:Area/Istex/Checkpoint">000F01</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">000F01</idno><idno type="wicri:doubleKey">1096-4959:1997:Bon E:plasma:vitellogenin:levels</idno><idno type="wicri:Area/Main/Merge">001949</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Plasma Vitellogenin Levels during the Annual Reproductive Cycle of the Female Rainbow Trout ( Oncorhynchus mykiss ): Establishment and Validation of an ELISA</title><author><name sortKey="Bon, E" sort="Bon, E" uniqKey="Bon E" first="E" last="Bon">E. 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Our results demonstrated that, under SDS-PAGE conditions, rainbow trout Vtg was composed of two molecular forms of 390 and 176 kDa representing, respectively, the dimeric form and the monomeric form of the molecule. The purified Vtg was used to raise a polyclonal antibody for Vtg (anti-Vtg). Using this anti-Vtg, a competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantification of rainbow trout Vtg. The practical sensitivity range of this ELISA was 20–320 ng/ml (80–20% of binding) and the detection limit was 9 ng/ml. The intra- and the inter-assay coefficients of variation (at 50% of binding) were estimated at 1.8% (n = 10) and 3.9% (n = 13), respectively. This ELISA was validated by detecting changes in Vtg levels in rainbow trout at different physiological stages, as well as in 2-year-old female rainbow trout throughout the reproductive cycle.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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