Competitive enzyme immunoassay for bovine growth hormone

Sang Gun Roh, Nobuyoshi Matsunaga, Akio Miyamoto, Satoshi Hidaka, Hisashi Hidari

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19 Citations (Scopus)

Abstract

We developed an enzyme immunoassay (A) for bovine GH (bGH) which is based on indirect competitive immunoassay in culture medium from a bovine pituitary cell culture. 40 μl 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 24 h at 37 °C. Biotin-label bGH was added and incubated further for 24 h at 37 °C, and biotinylated bGH was linked with streptoavidin-peroxidase. Substrates for peroxidase were added to the plate and incubated for 1 h at 4 °C. The enzyme reaction was stopped with 4N H2SO4, and the absorbency at 450 nm 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 x (EIA) - 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.

Original languageEnglish
Pages (from-to)195-198
Number of pages4
Journalendocrine journal
Volume44
Issue number1
DOIs
Publication statusPublished - 1997 Feb
Externally publishedYes

Keywords

  • Bovine GH
  • Enzyme immunoassay (EIA)
  • Radioimmunoassay (RIA)

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology

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  • Cite this

    Roh, S. G., Matsunaga, N., Miyamoto, A., Hidaka, S., & Hidari, H. (1997). Competitive enzyme immunoassay for bovine growth hormone. endocrine journal, 44(1), 195-198. https://doi.org/10.1507/endocrj.44.195