Newell D G, Bohane C, Payne S, Smith J L
J Immunol Methods. 1980;37(3-4):275-86. doi: 10.1016/0022-1759(80)90313-0.
A technique is described for the ultrastructural localisation of intracellular immunoglobulin (Ig) in human lymphoid cell suspensions by the immunoperoxidase method. The technique involves restricted saponin digestion of glutaraldehyde prefixed cells to enhance conjugate penetration. With this method of staining Ig was located in the rough endoplasmic reticulum, perinuclear space and Golgi apparatus in human lymphoid cells from a variety of sources, which is consistent with previously published observations using other ultrastructural techniques. In contrast, diffuse intracytoplasmic staining was predominant in cells prefixed with glutaraldehyde but not treated with saponin. These differences in patterns are discussed in terms of membrane permeability. Although saponin treatment was necessary for consistent localisation of intracellular Ig it resulted in unavoidable loss of Ig from the surface of the cells.
本文描述了一种通过免疫过氧化物酶法对人淋巴细胞悬液中细胞内免疫球蛋白（Ig）进行超微结构定位的技术。该技术包括对经戊二醛预固定的细胞进行有限的皂素消化，以增强偶联物的穿透。用这种染色方法，Ig位于来自各种来源的人淋巴细胞的粗面内质网、核周间隙和高尔基体中，这与以前使用其他超微结构技术发表的观察结果一致。相比之下，在经戊二醛预固定但未用皂素处理的细胞中，弥漫性胞质染色占主导。从膜通透性方面讨论了这些模式差异。虽然皂素处理对于细胞内Ig的一致定位是必要的，但它不可避免地导致细胞表面Ig的丢失。
