Characterization of monoclonal antibodies generated against bovine and porcine prostacyclin synthase and quantitation of bovine prostacyclin synthase

FEBS Letters

Ullrich, V.¹  Brugger, R.¹  Siegle, I.¹  Sprenger, R.¹  Nüsing, R.¹  Zecher, R.² 

[1] Faculty of Biology, PO Box 5560 (M612), University of Konstanz, 78434 Konstanz, Germany;Institute of Immunology, Obere Zahlbacher Straße 67, University of Mainz, Mainz, Germany

关键词: Monoclonal antibody;    Prostaglandin;    Prostacyclin synthase;    Immunohistochemistry;    Immunoquantitation;    ELISA;    enzyme-linked immunosorbent assay;    PGIS;    prostacyclin synthase;    PG;    prostaglandin;    HHD;    12-hydroxy-8;    10-heptadeca-dienoic acid;    PBS;    phosphate-buffered saline;    SDS-PAGE;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;   

DOI  :  10.1016/0014-5793(94)00504-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Monoclonal antibodies were raised against prostacyclin synthases purified from bovine and porcine aortae, respectively. Two monoclonal antibodies, RS1 and RS2, were purified and characterized. As shown by enzyme activity precipitation and Western blot analysis, in solubilized bovine and porcine aortae microsomes the monoclonal antibodies reacted only with prostacyclin synthase. The monoclonal antibody RS1 cross-reacts with partially purified prostacyclin synthase from human umbilical veins in an ELISA-based assay. None of the antibodies inhibited the enzyme activity. By combination of the monoclonal antibody RS2 with a polyclonal antibody we established an enzyme-linked immunosorbent assay (ELISA) for quantitation of bovine prostacyclin synthase. ELISA data were confirmed by Western blot analysis. Among different bovine tissues, aortae with 1665 ± 200 ng/mg microsomal protein showed the highest content of PGIS. Significant lower concentrations were observed in tongue, lung, kidney and thymus ranging from 49 ± 13.4 to 2.7 ± 0.9 ng/mg protein. The monoclonal antibody RS1 binds to endothelial cells and vascular smooth muscle cells in human liver tissue.

【 授权许可】

Unknown