Competitive enzyme immunoassay for bovine growth hormone.
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. 40 microliters 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 degrees C. Biotin-label bGH was added and incubated further for 24 h at 37 degrees C, and biotinylated bGH was linked with streptoavidin-peroxidase. Substrates for peroxidase were added to the plate and incubated for 1 h at 4 degrees 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 approximately 7.59% and 3.71 approximately 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.
Roh
S G
SG
Laboratory of Meat Animal Physiology, Obihiro University of Agriculture and Veterinary Medicine, Japan.
Matsunaga
N
N
Miyamoto
A
A
Hidaka
S
S
Hidari
H
H
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002940:Circadian Rhythm; D013006:Growth Hormone / Q000032:analysis* / Q000276:immunology; D007106:Immune Sera / Q000276:immunology; D007124:Immunoenzyme Techniques / Q000662:veterinary*; D010902:Pituitary Gland / Q000737:chemistry* / Q000166:cytology; D011817:Rabbits; D015203:Reproducibility of Results
D007106:Immune Sera; D013006:Growth Hormone