Schmidt W N, Page D L, Schmidt C A, McKusick K B, Ward W S, Hnilica L S
Cancer Res. 1984 Dec;44(12 Pt 1):5867-79.
Three stable monoclonal antibody-producing mouse hybridoma lines have been developed which produce high-titer, immunoglobulin M antibodies specific for the Novikoff ascites hepatoma (NAH) Mr 39,000 cytokeratin antigen (p39). Immunotransfer assays of cytoskeletal protein-enriched fractions indicated p39 to be present in a range of rat tissues, including colon, breast, lung, and uterus. Two-dimensional gel immunoblots confirmed that immunoreactivity in the latter tissues was for polypeptides with similar isoelectric points to those of NAH p39; however, reactivity in the colon contained a wide range of additional isomeric forms. Immunohistochemical localization studies with these antibodies revealed enrichment of p39 in the simple epithelia or ductular structures of organs containing this antigen. In all sections examined, glandular epithelia were observed to be only weakly immunoreactive. Additional immunoblot and immunocytochemical analyses with the monoclonal antibodies and polyspecific antisera to NAH cytokeratin suggest the human Mr 40,000 cytokeratin to be similar to but not identical to NAH p39.
已培育出三个稳定的产生单克隆抗体的小鼠杂交瘤细胞系，它们能产生高滴度的免疫球蛋白M抗体，该抗体对诺维科夫腹水肝癌（NAH）的39,000道尔顿细胞角蛋白抗原（p39）具有特异性。对富含细胞骨架蛋白的组分进行免疫印迹分析表明，p39存在于一系列大鼠组织中，包括结肠、乳腺、肺和子宫。二维凝胶免疫印迹证实，后几种组织中的免疫反应性针对的是等电点与NAH p39相似的多肽；然而，结肠中的反应性包含多种额外的异构体形式。用这些抗体进行的免疫组织化学定位研究显示，含有该抗原的器官的单层上皮或导管结构中p39富集。在所有检查的切片中，腺上皮的免疫反应性较弱。用针对NAH细胞角蛋白的单克隆抗体和多特异性抗血清进行的额外免疫印迹和免疫细胞化学分析表明，人40,000道尔顿的细胞角蛋白与NAH p39相似但不相同。
