Abstract
A monoclonal antibody (V4G6) to rat tissue kallikrein (EC 3.4.21.35) has been developed and characterized. This clone showed no cross-reactivity with rat tonin, rat esterase A or human urinary kallikrein in either radioimmunoassay or enzyme-linked immunosorbent assay. The monoclonal antibody used in the direct radioimmunoassay detects purified rat urinary kallikrein in a range of 0.32 to 40 ng per tube. The displacement curves for rat submandibular gland, pancreatic and kidney extracts and urine were parallel with the standard curve of purified rat urinary kallikrein. Analysis of immunoprecipitates from [14C] DFP labeled submandibular gland extract with SDS-polyacrylamide gel electrophoresis, demonstrates that this antibody recognizes only one 38,000 dalton serine protease while polyclonal antiserum identifies multiple species. Using this specific monoclonal reagent, tissue kallikrein was localized immunohistochemically in the granular convoluted tubules and striated ducts of the rat submandibular gland and in the acinar cells of the rat pancreas. The results showed that the monoclonal antibody (V4G6) can specifically recognize a single kallikrein in the tissue extracts without cross-reacting with other kallikrein-related serine proteases. This monoclonal antibody can be used as a specific reagent for quantitation, identification and immunohistochemical studies of tissue kallikrein.
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