Immunoreactivity of normal rabbit serum with epinephrine (E) cells of the rat adrenal medulla after microwave antigen retrieval.
Normal rabbit serum (NRS) produces intense staining of epinephrine (E) cells in microwave-heated sections of rat and mouse adrenal gland. This staining is not eliminated by liver adsorption, complement inactivation, high salt buffer, Triton X-100 or dilution in normal goat serum and bovine serum albumin (BSA), suggesting that it may result from specific antigen-antibody interactions. Western blots of adrenal medullary protein probed with NRS reveal several bands. The major band does not correspond to rat chromogranin A, which is a major constituent of E-cell secretory granules. The findings suggest that NRS may contain autoantibodies against a secreted rabbit E-cell protein with a homologous counterpart in rats and mice, and that this protein may be immunologically unmasked in situ by microwave heating. This phenomenon is a potential source of error in immunohistochemical studies of the adrenal medulla, and has potential biological significance in neuroimmunology.
Department of Pathology, Tufts University School of Medicine and New England Medical Center, 750 Washington Street, Box 802, Boston, MA 02111, USA. arthur.tischler@es.nemc.org
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Tischler
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Tsokas
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Shahsavari
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Powers
D000313:Adrenal Medulla / Q000166:cytology* / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000941:Antigens / Q000276:immunology / Q000528:radiation effects; D001323:Autoantibodies / Q000276:immunology; D015153:Blotting, Western; D019439:Chromaffin Cells / Q000378:metabolism; D004837:Epinephrine / Q000502:physiology*; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008872:Microwaves; D011817:Rabbits / Q000276:immunology; D051381:Rats; D011916:Rats, Inbred F344
D000941:Antigens; D001323:Autoantibodies; D004837:Epinephrine