Diethylpyrocarbonate, a vapour-phase fixative for immunofluorescence studies on polypeptide hormones

All the hormone antigens listed above react maximally with their antisera when the tissues containing them are freeze-dried and then fixed by diethylpyrocarbonate vapour, applied for 3 hr at 55°C.

Abstract

Immunofluorescence studies on polypeptide hormones require not only suitable antisera but also tissues in which the antigenic determinant sites of the hormones are free to react with antibody. Using antisera against ten hormones (gastrin, caerulein, secretin, enteroglucagon, insulin, glucagon, gastric inhibitory polypeptide, calcitonin, ACTH and growth hormone), we have tested a number of preparative procedures prior to indirect immunoflorescence. All the hormone antigens listed above react maximally with their antisera when the tissues containing them are freeze-dried and then fixed by diethylpyrocarbonate vapour, applied for 3 hr at 55°C. No other fixative or procedure has such universal applicability for immunofluorescence investigations on polypeptide hormones. For non-hormone proteins and polypeptides, the same may well be true.