Gown A M, Vogel A M, Gordon D, Lu P L
J Cell Biol. 1985 Mar;100(3):807-13. doi: 10.1083/jcb.100.3.807.
Injection of chicken gizzard actin into BALB/c mice resulted in the isolation of a smooth muscle-specific monoclonal antibody designated CGA7. When assayed on methanol-Carnoy's fixed, paraffin-embedded tissue, it bound to smooth muscle cells and myoepithelial cells, but failed to decorate striated muscle, endothelium, connective tissue, epithelium, or nerve. CGA7 recognized microfilament bundles in early passage cultures of rat aortic smooth muscle cells and human leiomyosarcoma cells but did not react with human fibroblasts. In Western blot experiments, CGA7 detected actin from chicken gizzard and monkey ileum, but not skeletal muscle or fibroblast actin. Immunoblots performed on two-dimensional gels demonstrated that CGA7 recognizes gamma-actin from chicken gizzard and alpha- and gamma-actin from rat colon muscularis. This antibody was an excellent tissue-specific smooth muscle marker.
将鸡胗肌动蛋白注射到BALB/c小鼠体内后,分离得到了一种平滑肌特异性单克隆抗体,命名为CGA7。在甲醇-卡诺氏固定、石蜡包埋的组织上进行检测时,它与平滑肌细胞和肌上皮细胞结合,但不能标记横纹肌、内皮细胞、结缔组织、上皮细胞或神经。CGA7可识别大鼠主动脉平滑肌细胞和人平滑肌肉瘤细胞早期传代培养物中的微丝束,但不与人成纤维细胞反应。在蛋白质印迹实验中,CGA7可检测到来自鸡胗和猴回肠的肌动蛋白,但不能检测到骨骼肌或成纤维细胞的肌动蛋白。在二维凝胶上进行的免疫印迹表明,CGA7可识别来自鸡胗的γ-肌动蛋白以及来自大鼠结肠肌层的α-和γ-肌动蛋白。该抗体是一种出色的组织特异性平滑肌标志物。