An enzyme-linked immunosorbent assay (ELISA) for measuring prolactin levels in ovine and cervine plasma

Abstract

Abstract An indirect enzyme-linked immunosorbent assay (ELISA) for ovine plasma prolactin that can be completed in less than 8 h is described and validated for ovine and cervine plasma. Ovine prolactin is covalently bound to bovine thyroglobulin and passively adsorbed in guanidine hydrochloride to a standard 96-well microtitre plate. Addition of standards, samples, and rabbit anti-ovine prolactin is specific for this assay, but subsequent steps which include addition of peroxidase-labelled goat-antirabbit IgG, washing, addition of o-phenylenediamine substrate with colour development, and reading of plates at 492 nm using an automatic ELISA processor are common to most ELISAs currently being performed in this laboratory. Assay sensitivity is less than 2.5 ng/ml, and intra- and inter-assay coefficients of variation are less than 9 and 16 % respectively. All steps except for sample addition are performed on a Behring Elisa Processor M automatic machine. This ELISA is quick, simple, and cost-effective compare...