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Solid‐Phase Immunoassay of PO Glycoprotein of Peripheral Nerve Myelin
Author(s) -
Nunn D. J.,
Mezei C.
Publication year - 1984
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1984.tb09712.x
Subject(s) - chemistry , antibody , immunoassay , glycoprotein , microbiology and biotechnology , peroxidase , antigen , chromatography , protein a , sodium dodecyl sulfate , myelin basic protein , myelin , biochemistry , biology , enzyme , immunology , central nervous system , neuroscience
To explore the immunological properties of PO protein, antibodies were elicited in rabbits against the purified chick PO protein. Peripheral nervous system protein was fractionated on sodium dodecyl sulfate‐polyacrylamide slab gels and then transferred electrophoretically (“blotted”) onto nitrocellulose sheets. The PO protein was detected by its capacity to bind its specific antibody present in the rabbit serum. The PO‐specific antibody complex was then exposed to goat anti‐rabbit immunoglobulin G (IgG) coupled to peroxidase or labeled with 125 I. The resulting PO antigen‐antibody “sandwich” was visualized and quantitated by densitometry of the colored peroxidase reaction product or by autoradiography and γ‐radiation counting of the 125 I‐IgG complex. The methods permitted quantitation of the PO protein in various nerve extracts. The limit of detection of the PO antigen was about 1 ng of protein. The antibody was specific for the PO glycoprotein in the peripheral nerve extracts. The PO proteins from various species, including human, were also detected by the antibody to chick PO protein. Preliminary experiments indicate the solid‐phase immunoassay is a useful method for monitoring PO protein levels in small quantities of tissue extracts under various physiological and pathological conditions.