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Development of a sandwich ELISA for determining plasma prolactin concentration in domestic birds

文献类型： 外文期刊

作者： Chen, R. 1 ; Guo, R. H. 1 ; Zhu, H. X. 1 ; Shi, Z. D. 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Anim Sci, Jiangsu Key Lab Food Qual & Safety, State Key Lab Cultivat Base,Minist Sci & Technol, Nanjing 210014, Jiangsu, Peoples R China

关键词： Prolactin; Goose; Sandwich; ELISA

期刊名称：DOMESTIC ANIMAL ENDOCRINOLOGY （ 影响因子：2.29； 五年影响因子：2.297 ）

ISSN： 0739-7240

年卷期： 2019 年 67 卷

页码：

收录情况： SCI

摘要： The present study was conducted to establish a sandwich ELISA for the determination of prolactin (PRL) concentrations in the plasma of domestic fowls. The assay uses a recombinant goose PRL as the reference standard, expressed in a eukaryotic system, and as the antigen for raising a polyclonal antibody in rabbit. This rabbit anti-goose PRL polyclonal antibody was used for coating the wells of the ELISA plate, and its biotinylated form served as the detection antibody. An avidin-conjugated horseradish peroxidase was used to bind the detection antibody and to catalyze the chromogenic reaction using 3,3',5,5'-tetramethylbenzidine as the substrate. The assay showed a linear relationship between the optical density and concentration of the standard PRL in the 0 to 12.5 ng/mL range, and the assay was sensitive to a concentration as low as 0.39 ng/mL. The intra- and inter-assay CVs were <7% and 11%, respectively. The response curves of the serially diluted plasma samples from goose, duck, and chicken exhibited similar parallel relationships to that observed for the reference standards. Consistent with previous findings, the assay effectively detected differences in PRL concentration in plasma samples from chicken, duck, and goose at various reproductive stages. (C) 2018 Elsevier Inc. All rights reserved.

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