go to jove.com

HIGH SCHOOL

Biology

Concept Videos

Cell Biology

This is a free preview. For full access

Visualizing Cells, Tissues, and Molecules

Antibody Staining in Cells and Tissues

Description

Immunocytochemistry (ICC) and immunohistochemistry (IHC) use antibodies to detect specific proteins or antigens in samples. ICC focuses on individual cells, such as blood cells and stem cells. IHC is used for tissue samples. These methods were first published by Albert Coons in 1941, when they were used to detect pneumococca...

Transcript

免疫细胞化学(ICC)和免疫组织化学(IHC)是利用酶联抗体来可视化和定量培养细胞或组织切片中特定蛋白质或抗原的染色技术。

在免疫细胞化学(ICC)中,将靶细胞单层培养在玻璃盖玻片上。用交联剂固定细胞以防止抗原被酶降解,然后用去垢剂处理使细胞膜通透。

在免疫组化(IHC)中,将整个组织固定并包埋在石蜡块中。然后将样品切成薄片,置于载玻片上。最后去除石蜡以恢复抗原。

当样本中的抗原暴露可及后,会与连接了偶联酶(如辣根过氧化物酶)的靶标特异性一抗结合。

该酶催化DAB染色剂氧化生成棕色沉淀,从而实现目标蛋白的可视化。

如果检测到的信号较弱,可使用带有偶联酶的二抗与靶蛋白上未标记的一抗结合,以进一步放大信号。