Dabelsteen E, Birkedal-Hansen H, Westergaard J, Fredebo L
J Invest Dermatol. 1980 Aug;75(2):136-9. doi: 10.1111/1523-1747.ep12521648.
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.
描述了一种用于分离质膜残留物的新方法，该方法可用于制备针对口腔上皮细胞膜的抗体。通过将汇合的大鼠口腔上皮细胞培养物原位暴露于100 mM新鲜制备的甲醛中，在细胞表面形成质膜囊泡。暴露后10 - 15分钟形成囊泡，并释放到培养基中。这些直径为1 - 10微米的囊泡通过在30,000 g下离心20分钟进行沉淀。在兔中制备针对囊泡的抗体，并将其用于间接免疫荧光和免疫过氧化物酶染色技术。在大鼠和人类口腔上皮中，它们可对基底细胞层和棘细胞层的细胞膜进行染色。在上皮中未见细胞质染色。对人子宫颈、大鼠和人类皮肤以及豚鼠唇部的鳞状上皮染色均为阴性。
