Journal of Animal Science and Technology

  • 제47권1호
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  • Pages.19-28
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  • 2005
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  • 2672-0191(pISSN)
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  • 2055-0391(eISSN)
한국축산학회 (Korean Society of Animal Sciences and Technology)
권호 표지
DOI QR코드

DOI QR Code

돼지 지방세포 원형질막 단백질에 대한 다클론항체의 생산 및 이용에 관한 연구

Studies on the Development and Utilization of Polyclonal Antibodies Against Swine Adipocyte Plasma Membrane Proteins

  • Baek, K.H. (National Livestock Research Institute, RDA) ;
  • Kwak, T.H. (Department of Animal science, Yeungnam University) ;
  • Oh, Y.S. (Institute of Biotechnology, Yeungnam University) ;
  • Choi, C.W. (National Livestock Research Institute, RDA) ;
  • Jung, K.K. (Department of Animal science, Yeungnam University) ;
  • Choi, Chang-Bon (Department of Animal science, Yeungnam University)
  • 발행 : 2005.02.28
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초록

The objectives of the current study were to develop polyclonal antibodies in sheep against adipocyte plasma membrane(APM) proteins isolated from swine, to investigate tissue specificity, and to determine cytotoxic effects of antiserum on swine adipocytes. Plasma membrane proteins from adipocyte, brain, heart, kidney, liver, and spleen were isolated using a 32% sucrose gradient. Adult male sheep was immunized three times at three week interval with the purified swine APM proteins. Antiserum was taken from immunized sheep at 10, 12, and 14 days after the third immunization. Antiserum expressed strong reactivity with APM proteins determined by enzyme-linked immunosorbent assay(ELISA), and the reactivity could be detected at dilutions in excess of 1 : 81,000. Antiserum showed very low binding affinity with proteins isolated from brain, heart, kidney, liver, or spleen. Tissue specificity of the antiserum was reconfirmed by Western immunoblotting using anti-sheep immunoglobulin G•alkalinephosphatase conjugate as a secondary antibody. The reactivity of antiserum to the external surface of fixed swine adipocytes was confmned by an immunohistochemical technique using anti-sheep immunoglobulin G-FITC. Confluent swine adipocytes in culture were lysed by antiserum treatment and cytosolie lactate dehydrogenase(LDH) was released as a dose-dependent patterns while adipocytes treated with normal sheep serum maintained their integrity and expressed low level of LDH. These results implicate that fat contents in the pigs can be reduced by immunological methods.

키워드

참고문헌

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  3. Development of Rapid Diagnostic Kit for Identification of Hanwoo (Korean Native Cattle) Brand Meat by Detecting BIO-TAG vol.34, pp.3, 2014, https://doi.org/10.5851/kosfa.2014.34.3.339