Lim S S, Tu Z H, Lemanski L F
J Muscle Res Cell Motil. 1984 Oct;5(5):515-26. doi: 10.1007/BF00713258.
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.
采用杂交瘤技术制备了抗肌钙蛋白 - T的单克隆抗体。最初使用酶联免疫吸附测定(ELISA)对培养上清液进行筛选。阳性克隆进行了两次亚克隆并进一步鉴定。其中之一,7/H3:C9:D10,产生了针对肌钙蛋白 - T的抗体;免疫印迹实验表明,当用多种横纹肌肌原纤维蛋白进行检测时,它仅对肌钙蛋白 - T具有特异性。用该抗体进行间接免疫荧光染色显示,在各种脊椎动物物种的成年和胚胎骨骼肌及心肌中均有特异性的I带染色。此外,在鸡胗平滑肌中可见强烈但弥漫的细胞质染色。我们的结果表明,肌钙蛋白 - T含有一种横纹肌和平滑肌共有的抗原决定簇。