The production of antibodies to cell membranes of squamous epithelium by the use of aldehyde-induced membrane vesicles.

A new method for the isolation of plasma membrane residues which can be used to raise antibodies to oral epithelial cell membranes is described. Plasma membrane vesicles were formed on the cell surfaces of confluent rat oral epithelial cell cultures by exposing the cells in situ to 100 mM freshly prepared formaldehyde. The vesicles formed 10--15 min. after exposure and were released into the medium. The vesicles, 1--10 micrometers in diameter, were sedimented by centrifugation at 30,000 g for 20 min. Antibodies to vesicles were raised in rabbits and used in an indirect immunofluorescence and immunoperoxidase staining technique. In rat and human oral epithelium they stained cell membranes in the basal and spinous cell layers. No cytoplasmic staining was seen in the epithelium. Staining of squamous epithelium from the human uterine cervix, rat and human skin and guinea pig lip was negative.