Journal of Histochemistry & Cytochemistry ( IF 3.2 ) Pub Date : 2021-09-19 , DOI: 10.1369/00221554211047287 Francesco Mascadri 1 , Maddalena M Bolognesi 1 , Daniela Pilla 2 , Giorgio Cattoretti 1, 2
Antigen-bearing proteins become progressively unavailable to immunodetection after prolonged storage of routine sections, exposed to a variety of agents, such as moisture, oxygen, and temperature. By proteomic analysis, the antigens are retained in the sections and definitely in the tissue block, pointing to fixation-independent, storage time–dependent protein modifications. Based on previous experience, we hypothesized that a combined exposure to a reducing agent and to chemicals favoring protein conformation changes would reverse the masking in aged sections. Disaccharides, lactose and sucrose, and a surfactant, added to a standard antigen retrieval buffer, reverse the negative changes in aged sections. Furthermore, they provide enhanced access to antigens in freshly cut sections, but not universally, revealing additional factors, besides heat and calcium chelation, required for antigen retrieval of individual proteins:
中文翻译:
恢复活力的老式组织切片在加工和长期储存过程中突出了个体抗原的命运
在常规切片长期储存、暴露于各种因素(如水分、氧气和温度)后,带有抗原的蛋白质逐渐无法用于免疫检测。通过蛋白质组学分析,抗原保留在切片中,并明确保留在组织块中,表明蛋白质修饰与固定无关,与储存时间有关。根据以前的经验,我们假设将还原剂和有利于蛋白质构象变化的化学物质联合暴露会逆转老化切片的掩蔽。添加到标准抗原修复缓冲液中的双糖、乳糖和蔗糖以及表面活性剂可逆转老化切片的负面变化。此外,它们提供了对新鲜切片中抗原的增强访问,但并非普遍适用,揭示了其他因素,