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Title	Development and characterization of monoclonal antibodies to rat tissue kallikrein and their use in biochemcal and immunohistochemical studies
Author	Woodley, Cheryl M.
Date	1984
College	College of Graduate Studies
Department	Molecular and Cellular Biology and Pathobiology
Extent	viii, 119 leaves; 29 cm
Subject.MeSH	Antibodies, Monoclonal Kallikreins--isolation & purification Immunoglobulin M Academic Dissertations
Collection	MUSC Theses and Dissertations
Contributing Institution	Waring Historical Library (MUSC)
Digitization Specifications	CanoScan 8400, Archival files are tiffs.
Date Digital	2015
Rights	Copyright is held by the author.
Language	English
SC County	Charleston County (S.C.)
Type	Text
Format	application/pdf
Media Type	Manuscripts
Resource Identifier	mtd1984_woodley_cm
Abstract	A panel of six mouse monoclonal antibodies (IgG1) against purified rat tissue kallikrein has been prepared based on the classical hybridoma technology. The clones were selected by their binding to kallikrein in both enzyme-linked immunosorbent assays and in direct radioimmunoassays. The antibodies were further characterized in regard to their binding titers, cross-reactivities, and their effect on enzyme activity. Binding titers giving 50% binding to 125I-kallikrein ranged from 1:2000 to 1:1,000,000. Antibodies from four of the clones showed no cross reactivity with human urinary kallikrein, rat urinary esterase A, or tonin using either assay. However, antibodies from a fifth clone cross reacted with rat tonin and from a sixth, with both rat tonin and urinary esterase A. Kallikrein activity studies indicate that two of the affinity purified antibodies inhibit activity, while one of the antibodies stimulates kallikrein activity. These monoclonal antibodies were used to identify kallikrein in tissue extracts from rat submaxillary gland, pancreas, kidney, and urine by either immunoprecipitation of [14C] DFP labeled proteins or Western blot analysis. In both assays, these antibodies bound a protein with Mr of 38,000 in SDS polyacrylamide gels. Analysis of in vitro translated poly A mRNA by immunoprecipitation with monoclonal antibodies revealed two kallikrein precursors with Mr's of 37,000 and 35,000 which were identified from submaxillary gland, pancreas, and kidney. The specificity of monoclonal reagents was further demonstrated in immunohistochemical localization of kallikrein in the granular convoluted tubules of the rat submaxillary gland and the acinar regions of the pancreas. Collectively, these data show that these monoclonal antibodies comprise a set of powerful and specific reagents for studies of tissue kallikrein.

Development and characterization of monoclonal antibodies to rat tissue kallikrein and their use in biochemcal and immunohistochemical studies

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