組換えタンパク質および合成ペプチドを抗原とした抗ウシレプチン抗体の作製とその特性について
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概要
- 論文の詳細を見る
The objective of this study was to generate antisera against recombinant bovine leptin and synthetic oligopeptides corresponding to the amino acid sequence 21-40 and 91-110 of bovine leptin. Recombinant bovine leptin was raised in the 293 cells and purified from 10 L of conditioned medium and utilized for immunization. The synthetic peptides were conjugated with keyhole limpet hemocyanin and inoculated into rabbits for antibody generation. Antibody titer was monitored by enzymeimmunoassay, immunoblotting and sandwich binding assay techniques. Each of the antisera, against three different antigens, was found to react with bovine leptin. The titers of anti-peptide antisera were lower than that of anti-recombinant leptin antiserum. Since anti-recombinant leptin antiserum was not neutralized by the leptin peptides 21-40 and 91-110, it is suggested that each antiserum recognizes a distinct epitope. In immunoblot analyses, all antisera exhibited cross-reactivity with human and mouse leptins. However, in the sandwich binding assay, the combination of anti-peptide antisera and anti-recombinant leptin antiserum, originated from bovine leptin, did not cross-react with either human or mouse leptin. The discrepancy of antigenic recognition between the immunoblot analyses and sandwich assay is thought to be dependent on the conformational status of leptin molecules between the species. The antisera generated in this study, which recognized distinct epitopes of bovine leptin, will provide a useful tool for studies of bovine leptin functions.
- 日本繁殖生物学会の論文
著者
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IMAI Kei
National Livestock Breeding Center
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高橋 透
National Institute of Agrobiological Sciences
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今井 敬
National Institute of Agrobiological Sciences
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橋爪 一善
Iwate University
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橋爪 一善
岩手大学農学部獣医学科
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TAKAHASHI Toru
National Institute of Agrobiological Sciences
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HASHIZUME Kazuyoshi
Iwate University
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