Isolation and characterization of a monoclonal antibody against the saxitoxin-binding component from the electric organ of the eel Electrophorus electricus
Abstract
A monoclonal hybridoma cell line secreting antibody against the saxitoxin-binding component from the eel Electrophorus electricus has been isolated. The specificity of this monoclonal antibody was established by (i) its ability to immunoprecipitate bound [3H]saxitoxin from a detergent extract of electroplax membranes in a dose-dependent manner, (ii) the inability of unrelated monoclonal antibodies to immunoprecipitate the toxin-binding activity in a similar assay, and (iii) the ability of excess unlabeled tetrodotoxin to displace [3H]saxitoxin from the immunoprecipitated component. The antibody is of the subclass IgG1 and binds specifically to a polypeptide component of Mr 250,000 on NaDodSO4/polyacrylamide gels. The antigenic determinant is associated with the same polypeptide component throughout the purification procedure, indicating that this component is not a result of artifactual aggregation or degradation during isolation. We conclude that the 250,000-dalton polypeptide is part of the saxitoxin binding/sodium channel protein in the native electroplax membrane.
Additional Information
© 1982 by the National Academy of Sciences. Communicated by Seymour Benzer, December 11, 1981. We thank David R. Balzer, Jr., for excellent assistance in the RIAs; Greg Lemke, Dr. Katherine A. Stygall, and Karl J. Fryxell for helpful discussions concerning hybridoma techniques and for gifts of control antibodies; Teresa Stevens for assistance in tissue culture; Dr. Raymond Withy for efforts in the preparation of [3H]STX; and Valerie Purvis for typing the manuscript and for the artwork. This research was supported by grants to M.A.R. and J.P.B. from the National Institutes of Health (NS 12018, NS 14403) and from the Pew Charitable Trust, a Postdoctoral Fellowship from the Muscular Dystrophy Association of America (to L.C.F.), and a Biomedical Research Grant (4 RR 0700 3A) from the National Institutes of Health to H.P.M. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.Files
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2008-02-11Created from EPrint's datestamp field
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2019-10-03Created from EPrint's last_modified field