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Immunological characterization of rat kininogens with monoclonal antibodies to T-kininogen.
- Source :
- European Journal of Biochemistry; 3/1/92, Vol. 204 Issue 2, p501-508, 8p
- Publication Year :
- 1992
-
Abstract
- A panel of 16 monoclonal antibodies (mAb) were produced against rat T-kininogen to characterize this family of proteins. These mAbs bound <superscript>125</superscript>I-T-kininogen by radioimmunoassay as well as reacting strongly with immobilized T-kininogen in an enzyme-linked immunosorbent assay (ELISA). The reactivity of these antibodies with proteolytic fragments of T-kininogen demonstrated the recognition of several different epitopes. One antibody was specific for the domain I of the heavy chain and/or the light chain, twelve antibodies were specific for domain 2 and three antibodies were specific for domain 3. All monoclonal antibodies recognized the two forms of T-kininogen encoded by the two different T-kininogen genes. T<subscript>1</subscript> and T<subscript>II</subscript> kininogen, except antibody TK 16-3.1 which uniquely reacted with T<subscript>II</subscript> kininogen. Two antibodies recognizing domain 2 cross-reacted with the high-molecular-mass kininogen (H-kininogen), whereas all the other monoclonal antibodies were specific to T-kininogen and did not recognize the heavy chain of H-kininogen. None of the antibodies tested altered the thiol protease inhibitory activity of T-kininogen, its partial proteolysis by rat mast cell chymase or the hydrolysis of H-kininogen by rat urinary kallikrein. The use of these antibodies in the development of sensitive ELISA to measure T-kininogen levels in plasma, urine, liver microsomes and hepatocytes is described. Two different forms of T-kininogen were distinguished by these monoclonal antibodies in Western blotting using rat plasma. The localization of T-kininogen was defined using these monoclonal antibodies by immunohistochemistry in rat liver hepatocytes and rat kidney. [ABSTRACT FROM AUTHOR]
- Subjects :
- MONOCLONAL antibodies
GLYCOPROTEINS
IMMUNOLOGY
BIOCHEMISTRY
IMMUNOHISTOCHEMISTRY
RATS
Subjects
Details
- Language :
- English
- ISSN :
- 00142956
- Volume :
- 204
- Issue :
- 2
- Database :
- Complementary Index
- Journal :
- European Journal of Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 13675033
- Full Text :
- https://doi.org/10.1111/j.1432-1033.1992.tb16660.x