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通过电子显微镜对弹性蛋白进行免疫定位

Immunologic localization of elastin by electron microscopy.

作者信息

Damiano V V, Tsang A, Christner P, Rosenbloom J, Weinbaum G

出版信息

Am J Pathol. 1979 Aug;96(2):439-56.

Abstract

Research on the pathogenesis of experimental emphysema has involved studies of the distribution of and destruction of elastin in the alveolar interstitium. The ill-defined organization of elastin in the alveolar interstitium makes it difficult to identify the elastin specifically by staining procedures ordinarily used for electron microscopy. This problem becomes more significant when the elastic tissue is fragmented during emphysema development and localization of the elastin fragments is essential. Therefore, a specific technique using high-titer antibodies against purified canine lung elastin was developed. The primary antibody was used on preembedded or etched postembedded sections. Localization of the antielastin IgG was accomplished with ferritin-labeled rabbit antisheep IgG as the secondary antibody. Treatment with the preimmune serum gave negligible ferritin background staining. The antielastin antibody did not react with lung connective tissue proteins such as the microfibrillar component of elastin or collagen or proteoglycan. The antielastin antibody appeared to be species specific. The method may be useful for studies of experimental emphysema.

摘要

实验性肺气肿发病机制的研究涉及对肺泡间质中弹性蛋白分布及破坏情况的研究。肺泡间质中弹性蛋白的组织结构不明确,使得通过通常用于电子显微镜检查的染色程序特异性识别弹性蛋白变得困难。当弹性组织在肺气肿发展过程中发生碎片化且弹性蛋白片段的定位至关重要时,这个问题就变得更加突出。因此,开发了一种使用针对纯化犬肺弹性蛋白的高滴度抗体的特异性技术。将一抗用于预包埋或蚀刻后的包埋切片。以铁蛋白标记的兔抗羊IgG作为二抗来完成抗弹性蛋白IgG的定位。用免疫前血清处理产生的铁蛋白背景染色可忽略不计。抗弹性蛋白抗体不与肺结缔组织蛋白如弹性蛋白的微纤维成分、胶原蛋白或蛋白聚糖发生反应。抗弹性蛋白抗体似乎具有种属特异性。该方法可能对实验性肺气肿的研究有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bfa/2042436/5b8879cdef10/amjpathol00240-0098-a.jpg

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