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dc.contributor.authorMaioli, Marcos Antonio [UNESP]
dc.contributor.authorNogueira, Guilherme de Paula [UNESP]
dc.date.accessioned2018-12-11T17:36:22Z
dc.date.available2018-12-11T17:36:22Z
dc.date.issued2017-12-01
dc.identifierhttp://dx.doi.org/10.1590/S0100-736X2017001200030
dc.identifier.citationPesquisa Veterinaria Brasileira, v. 37, n. 12, p. 1545-1553, 2017.
dc.identifier.issn1678-5150
dc.identifier.issn0100-736X
dc.identifier.urihttp://hdl.handle.net/11449/179688
dc.description.abstractThis study aimed to standardize an in house competitive enzyme-linked immunosorbent assay (cELISA) to determine plasma concentrations of total insulin-like growth factor I (IGF-I) for the bovine specie using the amplification biotin-streptavidin peroxidase system. The IGF-I was extracted from insulin-like growth factor binding proteins (IGFBPs) using the acidified glycine buffer followed by the pH neutralization with sodium hydroxide. The microplates were coated with anti-rabbit IgG, thereafter the measurements were carried out using two approaches, one with a prior incubation of samples with the anti-h-IGF-I antibody and another without previous incubation (simultaneous addition of IGF-I and biotinylated sample). The best results were obtained using the method without the prior incubation, using the following combination of reagents: microplates were coated with 0.25μg/well of anti- rabbit IgG, the specific antibody at a dilution of 1:250,000 and 0.06ng/well of biotinylated IGF-I. The in house methodology showed sensitivity of 50ng/ml, a correlation between doses of 0.945 when compared to a commercial method. In addition, after 33 assays (quantification of 1114 samples) the proposed methodology presented a good precision, with inter-assay variation coefficients of 12.94% and 20.71% for the high and low controls, respectively. Finally, we concluded that ELISA method for the quantification of total IGF-I using the system biotin-streptavidin-peroxidase amplification in a competitive assay is established and is presented as a useful tool for studies aimed at monitoring the IGF-I concentrations.en
dc.format.extent1545-1553
dc.language.isopor
dc.relation.ispartofPesquisa Veterinaria Brasileira
dc.sourceScopus
dc.subjectBovine
dc.subjectELISA
dc.subjectHormone
dc.subjectIGF-I
dc.subjectInsulin-like growth factor I
dc.subjectPlasm
dc.subjectValidation
dc.titlePadronização da quantificação do fator de crescimento semelhante à insulina I (IGF-I) em plasma bovino por ELISA1pt
dc.title.alternativeStandardization of ELISA for the measurement of Insulin-Like Growth Factor I (IGF-I) in bovine plasmen
dc.typeArtigo
dc.contributor.institutionUniversidade Estadual Paulista (UNESP)
dc.description.affiliationDepartamento de Apoio a Produção e Saúde Animal Faculdade de Medicina Veterinária Universidade Estadual Paulista Júlio de Mesquita Filho (Unesp), Rua Clóvis Pestana 793, Dona Amélia
dc.description.affiliationUnespDepartamento de Apoio a Produção e Saúde Animal Faculdade de Medicina Veterinária Universidade Estadual Paulista Júlio de Mesquita Filho (Unesp), Rua Clóvis Pestana 793, Dona Amélia
dc.identifier.doi10.1590/S0100-736X2017001200030
dc.identifier.scieloS0100-736X2017001201545
dc.rights.accessRightsAcesso aberto
dc.identifier.scopus2-s2.0-85044202650
dc.identifier.fileS0100-736X2017001201545.pdf
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