Developing an improved competitive enzyme immunoassay for beetle caprine growth hormone
2011
Gul, R. (Punjab Univ., Lahore (Pakistan). School of Biological Sciences) | Ghafoor, F. (Sheikh Zayed Medical Complex, Lahore (Pakistan). NHRC) | Sadaf, S. (Inst. of Biochemistry and Biotechnology, Lahore (Pakistan)) | Akhtar, M.W.
The study aimed to develop a simple and sensitive competitive enzyme immunoassay for growth hormone (GH) in Beetle caprine serum. Antisera (rabbit anti-rcGH) were raised in two rabbits of local breed. Periodate method was used for labeling horseradish peroxidase (HRP) with the purified recombinant caprine growth hormone (rcGH). 96-well microtitre plates were coated with primary antibody (polyclonal rabbit anti-rcGH) for 24 hr at 4 degree C. Buffer standards ranging from 0-400 ng/ml were prepared with rcGH to be used in each assay batch. Assay parameters like pH, temperature, reaction time and working dilutions of antibody & conjugate were optimized. The assay showed in significant cross-reactivity with the human insulin. The developed assay is reliable, quick and simple to perform.
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