Selective but nonspecific immunolabeling of enamel protein-associated compartments by a monoclonal antibody against vimentin
Selective but nonspecific immunolabeling of enamel protein-associated compartments by a monoclonal antibody against vimentin
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DOI：
10.1177/002215549904701003
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发表时间：
1999-10-01
影响因子：
3.2
通讯作者：
Nanci, A
中科院分区：
文献类型：
--
作者：
Josephsen, K;Smith, CE;Nanci, A
波形蛋白，一种中间丝成分，已被确定在许多间充质细胞的各种LM和EM免疫标记技术。在我们的研究中，几种组织处理条件和波形蛋白的单克隆和多克隆抗体进行筛选大鼠切牙成牙本质细胞的免疫染色。使用植入后的胶体金免疫细胞化学，我们无法检测到任何令人信服的波形蛋白抗原在这些细胞中，但单克隆抗体（V9-S）出乎意料地导致强烈的标记内和细胞外室，通常是强烈的免疫反应性与抗釉原蛋白抗体。阻断实验表明，V9-S结合被抗釉原蛋白抗体竞争。免疫印迹表明，釉质蛋白反应后，用戊二醛固定与抗波形蛋白抗体。这些数据表明，所观察到的免疫反应是针对一个表位显然是由釉质蛋白在固定过程中交联。虽然标记不能被认为是特异性的，但它仍然是选择性的，因为它非常精确地定位在含有釉质蛋白的隔室上，并且不与其他钙化的牙齿组织（包括牙本质和骨）结合。因此，V9-S抗体可用作鉴定固定组织中釉原蛋白的存在和分布的可靠探针。
Vimentin, an intermediate filament component, has been identified in many mesenchymal cells by a variety of LM and EM immunolabeling techniques. In our study, several tissue-processing conditions and monoclonal and polyclonal antibodies against vimentin were screened for immunostaining of rat incisor odontoblasts. Using postembedding colloidal gold immunocytochemistry, we were unable to detect any convincing vimentin antigenicity in these cells, but one of the monoclonal antibodies (V9-S) unexpectedly resulted in intense labeling over intra- and extracellular compartments that normally are strongly immunoreactive with anti-amelogenin antibodies. Blocking experiments showed that V9-S binding was competed by anti-amelogenin antibody. Immunoblots indicated that enamel proteins reacted with this anti-vimentin antibody after fixation with glutaraldehyde. These data suggest that the observed immunoreaction is directed against an epitope apparently created by crosslinking of enamel proteins during fixation. Although the labeling cannot be considered specific, it is nevertheless selective because it is very precisely localized over compartments containing enamel proteins and shows no binding to other calcified dental tissues, including dentin and bone. The V9-S antibody can therefore be used as a reliable probe to identify the presence and distribution of amelogenins in fixed tissues.