Production and characterization of a monoclonal antibody directed against the 43,000-dalton v1 polypeptide from Torpedo marmorata electric organ.

摘要

Subsynaptic membrane fragments prepared from Torpedo marmorata electric organ contain, in addition to the acetylcholine receptor polypeptides, a major protein band of apparent molecular mass 43,000 daltons. On two-dimensional gels, this band yields three spots referred to as v1, v2, and v3. Monoclonal antibodies against the 43,000-dalton proteins were developed in CBA mice. One of them reacted exclusively with the v1 polypeptide but not with v2 and v3. Staining by the "immunogold" reaction followed by observation by electron microscopy showed that this antibody exclusively labeled the innervated membrane of T. marmorata electroplaque on its cytoplasmic face. Electroblots of one-dimensional gels of membrane preparations from 80-mm embryo electric organ were prepared. After reaction with the anti-v1 monoclonal antibody, a strongly stained 43,000-dalton band was revealed.