| Publication number |
CLPUB00400 |
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| Authors |
Nishikawa T., Maruo N., Tatarazako N., Shiraishi H., Morita M. |
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| Title |
Establishment and characterization of medaka vitellogenin monoclonal antibodies. |
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| Citation |
(In) 4th annual meeting of Japan Society of Endocrine Disrupters Research; pp.PB-61; Tsukuba (2001) |
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| Web pages |
https://www.env.go.jp/chemi/end/sympo/2001/report/pdf_abs/pb/PB_61.pdf |
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| Abstract |
We established Medaka (Oryzias latipes) vitellogenin (VTG) monoclonal
antibodies for ELISA application. Medaka Was fed by 1 mg 17 beta-
estradiol/g tetrafin for seven days to collect ascite fluids containing
VTG. After purification of VTG using POROS-HQ column chromatography, VTG
in PBS was mixed with equal volume of Freund's adjuvant and Mice (6 weeks
BALB/c females) were immunized 3 times per every 14 days. Spleen cells
from immunized mouse and myeloma cells were fused using polyethylene
glycol, then hybridoma cells were subjected to HAT selection. Hybridoma
producing monoclonal antibodies specific to Medaka VTG were selected and
four clones (MVP25, MVP47, MVP49, MVP51) were established, of which these
four monoclonal antibodies did not show any reactivity to male blood and
whole body and liver homogenates. Western blot analysis revealed that each
of these antibodies recognized lipovitellin heavy chain (120 kDa protein
existed in VTG N-terminal region) as an epitope. It was possible to
construct sandwich ELISA system for Medaka VTG using MVP25-MVP47 or MVP47-
MVP49, but not MVP25-MVP49, indicating MVP25 and MVP49 bind the same or
near epitope.
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| Cell lines |
CVCL_U131; MVP25 CVCL_U132; MVP47 CVCL_U133; MVP49 CVCL_U134; MVP51 |
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