Anti-troponin-T monoclonal antibody crossreacts with all muscle types

Abstract

Monoclonal antibodies to troponin-T were produced by the hybridoma technique. Culture supernatants were initially screened using an enzyme-linked immunoabsorbent assay (ELISA). Positive clones were subcloned twice and further characterized. One of these, 7/H3:C9:D10, produced antibodies against troponin-T; immunoblotting experiments indicated its specificity for only troponin-T when challenged with a variety of striated muscle myofibrillar proteins. Indirect immunofluorescence staining with the antibody shows specific I-band staining in both adult and embryonic skeletal and cardiac muscle of various vertebrate species. In addition, intense but diffuse cytoplasmic staining was seen in chicken gizzard smooth muscle. Our results suggest that troponin-T contains an antigenic determinant that is common to both striated and smooth muscle.