Competitive Enzyme Immunoassay for Bovine Growth Hormone

Endocrine Journal

SATOSHI HIDAKA¹  NOBUYOSHI MATSUNAGA¹  HISASHI HIDARI¹  AKIO MIYAMOTO²  SANG-GUN ROH¹ 

[1] Laboratory of Meat Animal Physiology, Department of Animal Science, Obihiro University of Agriculture and Veterinary Medicine;Laboratory of Animal Genetics and Reproduction, Department of Animal Science, Obihiro University of Agriculture and Veterinary Medicine

关键词: Enzyme immunoassay (ETA);    Radioimmunoassay (RIA);    Bovine GH;   

DOI  :  10.1507/endocrj.44.195

学科分类：内分泌与代谢学

来源: Japan Endocrine Society

【 摘 要 】

References(6)Cited-By(12)We developed an enzyme immunoassay (EIA) for bovine GH (bGH) which is based on indirect competitive immunoassay in culture medium from a bovine pituitary cell culture. 40μl cell culture samples (or bGH standard) and bGH antibody (rabbit anti-bGH) were added to the 96 well microplate coated with secondary antibody (Goat anti-rabbit IgG), and incubated for 24h at 37°C. Biotin-label bGH was added and incubated further for 24h at 37°C, and biotinylated bGH was linked with streptoavidin-peroxidase. Substrates for peroxidase were added to the plate and incubated for 1h at 4°C. The enzyme reaction was stopped with 4N H2SO4, and the absorbency at 450nm was measured with an ELISA Reader. The coefficients of intra-assay and inter-assay variations were 4.13-7.59% and 3.71-8.27%, respectively. The regression equation and correlation coefficients with the radioimmunoassay (RIA) were y(RIA)=1.9986×(ETA)-1.3921 and 0.9701(n=27), respectively. Collectively, the present assay provides a reliable alternative to RIA and offers the major advantage of eliminating radioactive reagents and counting equipment.

【 授权许可】

Unknown