Demonstration of the display of components of the endoplasmic reticulum system by indirect immunofluorescence microscopy using antibodies against cytochrome b(5) from rat liver microsomes.

Antibodies against the purified fragment of cytochrome b(5) released by trypsin treatment from rat liver microsomes were raised in rabbits and used for the demonstration of membranes rich in cytochrome b(5), in particular the endoplasmic reticulum (ER) system, by indirect immunofluorescence microscopy. The method allowed the demonstration of ER not only in frozen sections of various tissues, including liver and lactating mammary gland from different species, but also in cultured cells of a diversity of species and cell types. In the cultured cells the structures most prominently decorated with the antibodies against cytochrome b(5) were the nuclear envelope and the ER system which in many cells could be recognized as a system of smoothly bending, branching threads, extending from the perinuclear cytoplasm toward the cell periphery. Changes in the display of such elements during mitosis and cell plating and possible influences of the specific fixations used are discussed.