The use of detergents and immunoperoxidase reagents for the ultrastructural demonstration of internal immunoglobulin in lymph cells.
Lymph-borne immunoblasts were fixed in dilute glutaraldehyde and then treated with saponin. This treatment made most parts of the cells permeable to ferritin, so that anti-immunoglobulin (Ig) antibodies which had been conjugated to horse radish peroxidase (HRP) had no difficulty in gaining access to Ig which thus could be demonstrated at an ultrastructural level. Best results were obtained by fixing the cells in 0.1% glutaraldehyde for 7 min and then treating them with a 1% solution of saponin for 100 min at 55 degrees C before exposing them to the Ig-HRP conjugate. The method yielded reproducible results and although it causes a small amount of ultrastructural damage, it may be of value in detecting a variety of intracellular antigens.
D000818:Animals; D003902:Detergents; D005404:Fixatives; D007124:Immunoenzyme Techniques; D007136:Immunoglobulins / Q000378:metabolism*; D008196:Lymph / Q000276:immunology*; D008214:Lymphocytes / Q000276:immunology* / Q000648:ultrastructure; D012503:Saponins; D012756:Sheep
D003902:Detergents; D005404:Fixatives; D007136:Immunoglobulins; D012503:Saponins