Synopsis
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.
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References
Anfinsen, C. B. (1973). Principles that govern folding of protein chains.Science 182, 223–30.
Coons, A. H., Leduc, E. H. &Connolly, J. M. (1955). Studies on antibody production. I. A method for the histochemical demonstration of specific antibody and its application to a study of the hyperimmune rabbit.J. exp. Med. 102, 49–60.
Kendall, P. A., Polak, J. M. &Pearse, A. G. E. (1971). Carbodiimide fixation for immunohistochemistry: Observations on the fixation of polypeptide hormones.Experientia 27, 1104–6.
Polak, J. M., Bloom, S., Coulling, I. &Pearse, A. G. E. (1971). Immunofluorescent localization of enteroglucagon cells in the gastrointestinal tract of the dog.Gut 12, 311–18.
Starke, G. R. (1970). Recent developments in chemical modification and sequential degradation of proteins.Adv. Prot. Chem. 24, 261–308.
Thoma, W. &Rinke, H. (1959). Syntheses with pyrocarbonic esters.Liebigs Ann. Chem. 624, 30–6.
Wolf, B., Lesnaw, J. A. &Reichmann, M. E. (1970). A mechanism of the irreversible inactivation of bovine pancreatic ribonuclease by diethylpyrocarbonate.Eur. J. Biochem. 13, 519–25.
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Pearse, A.G.E., Polak, J.M., Adams, C. et al. Diethylpyrocarbonate, a vapour-phase fixative for immunofluorescence studies on polypeptide hormones. Histochem J 6, 347–352 (1974). https://doi.org/10.1007/BF01312253
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DOI: https://doi.org/10.1007/BF01312253