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||2008 Japanes Assay kit Catalog-p839
||In recent years, influence of Endocrine Disrupting Chemicals (EDC) upon living organisms has been one of the hot topics in the environmental science. However, evaluation methods of chemical compound species that may have EDC effects has not yet been established, and it is absolutely necessary to take action to complete such a task. Vitellogenin (Vg), a precursor of yolk protein, is a female-specific protein that is present only in the blood of higher oviparous vertebrate organisms. Vitellogenin is normally expressed in the liver under regulation of estrogen (female sex hormone) during an egg formation process, and when it reaches a growing oarian follicle via blood stream it contributes to the egg yolk formation. Since estrogen is known to have an effect upon male species to induce unusual expression of Vg, an ELISA screening method detecting Vg in EDC exposed male animals is the most reliable bio-screening assay system.
1. Antibody-coated microtiter plate (8 wells ~12) : 1 plate
2. The standard, Japanese Quail Vg, lyophilized : 300 ul ~ 2
3. Dilution solution : 30 mL ~ 2
4. Anti-Japanese Quail Lipovitellin polyclonal antibody (~100) : 80 ul ~ 1
5. HRP-conjugated anti-Rabbit IgG (~100) : 80 ul ~ 1
6. OPD (o-phenylendiamine) tablets : 2 tab
7. Substrate buffer : 15 mL ~ 1
8. Stop solution : 6 mL ~ 1
9. Wash solution (~20) : 30 mL ~ 1
Dynamic range : 7.8 - 200 ng / mL
This kit is based on the ELISA method which uses two different specific anti-Japanese Quail Vg antibodies. Anti-Japanese Quail monoclonal antibody is coated on the microplate, so that Vg in the sample and standard solution is captured. Reacting with anti-Japanese Quail polyclonal antibody and HRP-conjugated anti-Rabbit IgG, the complex of ''coated-antibody - antigen - antibody'' is formed. Finally, Vg concentrations are estimated according to the coloring intensity developed with enzymatic reaction.
• The Effect of Estrogens, River Water, and Treated Wastewater on Vitellogenin Induction of Japanese Medaka Kiyoaki Kitamura, Kiyoshi Miyajima, Tadashi Higashitani, Norihide Nakada ,Koya Komori, Yutaka Suzuki, JAWRA, Volume 45, Issue 1, pages 22-34, February 2009
• Reproductive effects and bioconcentration of 4-nonylphenol in medaka fish (Oryzias latipes). Ishibashi H, Hirano M, Matsumura N, Watanabe N, Takao Y, Arizono K., Chemosphere. 2006 Nov;65(6):1019-26.
• Effects of pentachlorophenol on the reproduction of Japanese medaka (Oryzias latipes). Zha J, Wang Z, Schlenk D., Chem Biol Interact. 2006 May 15;161(1):26-36.
• Development of on-site fish exposure system placed in water quality monitoring stations along a river. Higashitani T, Miyajima K, Nakada N, Yasojima M, Tanaka H, Suzuki Y., Water Sci Technol. 2005;52(12):275-82.
• Validation of an Enzyme-Linked Immunosorbent Assay Method for Vitellogenin in the Medaka Norihisa Tatarazako, Masaaki Koshio, Hiroyuki Hori, Masatoshi Morita and Taisen Iguchi, JOURNAL OF HEALTH SCIENCE Vol. 50 (2004) , No. 3 301-308
• Fish full life-cycle testing for androgen methyltestosterone on medaka (Oryzias latipes). Seki M, Yokota H, Matsubara H, Maeda M, Tadokoro H, Kobayashi K., Environ Toxicol Chem. 2004 Mar;23(3):774-81.
• Effects of triclosan on the early life stages and reproduction of medaka Oryzias latipes and induction of hepatic vitellogenin. Ishibashi H, Matsumura N, Hirano M, Matsuoka M, Shiratsuchi H, Ishibashi Y, Takao Y, Arizono K., Aquat Toxicol. 2004 Apr 14;67(2):167-79.
• Effects of 4-nonylphenol and 4-tert-octylphenol on sex differentiation and vitellogenin induction in medaka (Oryzias latipes). Seki M, Yokota H, Maeda M, Tadokoro H, Kobayashi K. Environ Toxicol Chem. 2003 Jul;22(7):1507-16.