Blaustein D I, Dewey M M
J Histochem Cytochem. 1979 Mar;27(3):788-93. doi: 10.1177/27.3.314460.
Localization of rhodopsin and its position in the membrane has been the subject of numerous studies. Most recently, immunocytochemical techniques have been employed to localize the opsin component of the molecule in in situ rod outer segments. Due to the problems inherent in localization procedures (penetration and mechanical interference) we have utilized isolated, osmotically intact rod outer segment discs in this study. Specific antibodies to chromatographically pure rhodopsin were prepared and enzymatically digested to their Fab components. The univalent Fab antibodies were conjugated to horseradish peroxidase and used to label the isolated rod outer segment discs. Discs treated with anti-opsin conjugate stained uniformly and heavily on their interdisc surfaces. Reaction product was also present on the intradisc surface in a thinner but still uniformly distributed layer. Controls treated with preimmune Fab - horseradish peroxidase conjugate showed no deposition of reaction product.
视紫红质的定位及其在膜中的位置一直是众多研究的主题。最近，免疫细胞化学技术已被用于在原位视杆细胞外段中定位该分子的视蛋白成分。由于定位程序中固有的问题（穿透和机械干扰），我们在本研究中使用了分离的、渗透压完整的视杆细胞外段盘。制备了针对色谱纯视紫红质的特异性抗体，并将其酶解为Fab片段。单价Fab抗体与辣根过氧化物酶偶联，并用于标记分离的视杆细胞外段盘。用抗视蛋白偶联物处理的盘在其盘间表面均匀且强烈地染色。反应产物也以较薄但仍均匀分布的层存在于盘内表面。用免疫前Fab-辣根过氧化物酶偶联物处理的对照未显示反应产物的沉积。
