An attempt of the development of solid-phase enzyme immunoassay of gonadotropin releasing hormone.
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- GESHI Masaya
- Department of Animal Reproduction, College of Agriculture, University of Osaka Prefecture Tohoku National Agricultural Experiment Station
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- KASUGA Naoki
- Research Division, Otsuka Assay Laboratory, Otsuka Pharmaceutical Co., Ltd.
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- IMAGAWA Kenichi
- Research Division, Otsuka Assay Laboratory, Otsuka Pharmaceutical Co., Ltd.
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- MORI Junichi
- Department of Animal Reproduction, College of Agriculture, University of Osaka Prefecture
Bibliographic Information
- Other Title
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- 性腺刺激ホルモン放出ホルモンの固相法Enzyme Immunoassay法に関する検討
- セイセン シゲキ ホルモン ホウシュツ ホルモン ノ コソウホウ enzyme
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Abstract
A solid-phase competitive enzyme immunoassay of gonadotropin releasing hormone (GnRH) was examined. Enzyme-labeled antigens were prepared by coupling synthetic GnRH with peroxidase using bis-diazotized benzidine and purified by Sephadex G-25 column chromatography. Antiserum raised against [Glu1]-GnRH-rabbit serum albumin conjugate in a male horse was used. Immunoglobulin G (IgG) fraction of the antiserum used for a solid-phase was prepared by an (NH4)2SO4 precipitation method, purified by Sephacryl S-300 column chromatography, and adsorbed to a polystyrene ball.<BR>Three-tenths ml of 0.05 M phosphate buffer (pH 7.5), 0.1 ml of standard or sample solution, 0.1 ml of peroxidase-labeled GnRH solution and a solid-phase polystyrene ball were added in a test tube. After incubation at 4°C for 24 hours, a polystyrene ball was washed three times with 2 ml of saline, and transferred to a new tube. The enzyme activity was measured with a spectrophotometer (absorbance at 492 nm).<BR>A dose-response curve was obtained between 1 to 1000 ng/ml of GnRH when a solid-phase coated with 50 μg/ml IgG solution and peroxidase-labeled GnRH solution (×100) were used. Acid extract of bovine pituitary stalk produced dose-response curve indistinguishable from synthetic GnRH, but those of anterior lobe, liver and muscle didn't produced.<BR>These results show that GnRH is assayed by this method.
Journal
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- The Japanese journal of animal reproduction
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The Japanese journal of animal reproduction 32 (4), 172-176, 1986
THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT
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Details 詳細情報について
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- CRID
- 1390001206333689216
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- NII Article ID
- 130004214131
- 40004157299
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- NII Book ID
- AN00041843
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- NDL BIB ID
- 3122579
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- ISSN
- 03859932
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- Text Lang
- ja
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- Data Source
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- JaLC
- NDL
- Crossref
- CiNii Articles
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- Abstract License Flag
- Disallowed