Distribution of Murine Tissue Specific ${\gamma}$-Glutamyltransferase: -Comparison of Six Monoclonal Antibody Applications in Enzyme Linked Immunosorbent Assay, Radioimmunoassay, Immunohistochemistry, and Autoradiography-

${\gamma}$-Glutamyltransferase의 조직내 분포에 관한 연구 -단일클론항체의 효소면역측정법, 방사면역측정법, 면역조직화학검사, 자가방사기록검사 적용에 관하여 -

  • Kim, Meyoung-Kon (Department of Biochemistry, College of Medicine, Korea University) ;
  • Park, Youn-Kyu (Department of Biochemistry, College of Medicine, Korea University) ;
  • Ryu, Chong-Kun (Department of Biochemistry, College of Medicine, Korea University)
  • 김명근 (고려대학교 의과대학 생화학교실) ;
  • 박윤규 (고려대학교 의과대학 생화학교실) ;
  • 류총근 (고려대학교 의과대학 생화학교실)
  • Published : 1994.05.15

Abstract

${\gamma}$-Glutamyltransferase (GGT: E.C. 2.3.2.2.) is a glycoprotein enzyme which is involved in glutathione metabolism and amino acid transport through the plasma membrane. It is distributed widely in several organs including liver and kidney. Several isozymes of GGT have been reported and some of the isozymes may be associated with hepatocarcinogenesis. We have produced six monoclnal antibodies (mAbs) against GGT purified from the liver of 2-acetamidofluorene (AAF) treated rats. All of the six mAbs were obtained by immunizing mice with liver GGT Six hybridomas which produced anti-GGT Abs were extensively subcloned and injected into the peritoneal cavity of BALB/c mice to obtain large quantities of Abs. These mAbs were purified from ascites by ammonium sulfate precipitation and protein A sepharose CL-4B column chromatography. Using these mAbs we preformed enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunohistochemistry (IHC), and autoradiography (ARG) to study the distribution of GGT isozyme in tissue. The results indicate that GGT-mAb 1 is specific for the AAF treated liver GGT, GGT-mAb 5 for the normal liver GGT, and GGT-mAb 6 for the normal kindey GGT. These mAbs may be used to evaluate the distribution of GGT isozymes in different tissues.

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