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March 1, 1982Proceedings of the National Academy of SciencesOpen Access

Isolation and characterization of a monoclonal antibody against the saxitoxin-binding component from the electric organ of the eel Electrophorus electricus.

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Authors

HMH P MooreFlorida State UniversityLFL C FritzGerman Cancer Research CenterMRMichael A. RafteryUniversity of Notre Dame

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Implication

In vitro study reveals a monoclonal antibody targeting a 250-kilodalton polypeptide in electric eel membranes, indicating this component constitutes the native voltage-gated sodium channel.

Key Points

  • Isolate and characterize a monoclonal antibody specific to the saxitoxin-binding component of the sodium channel from Electrophorus electricus electroplax tissue.
  • Isolated a monoclonal hybridoma cell line producing an IgG1 antibody against the saxitoxin-binding component from electric eel electroplax membranes.
  • Evaluated antibody specificity using dose-dependent immunoprecipitation of [3H]saxitoxin and competitive displacement with excess unlabeled tetrodotoxin.
  • Analyzed the molecular weight and stability of the target polypeptide across isolation stages using sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
  • The isolated IgG1 antibody selectively immunoprecipitated membrane-bound [3H]saxitoxin in a dose-dependent manner, whereas control antibodies showed no activity.
  • Unlabeled tetrodotoxin successfully displaced bound [3H]saxitoxin from the immunoprecipitated component.
  • The antibody bound specifically to a 250,000-dalton polypeptide across all purification steps, verifying that this protein is an authentic component of the native sodium channel rather than an artifact.

Cite This Study

Moore et al. (1982) studied this question.

synapsesocial.com/papers/6a95ca615ec4b71689c5d5a1https://doi.org/10.1073/pnas.79.5.1673
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Also Consider

Synapse has enriched 4 closely related papers on similar clinical questions. Consider them for comparative context:

  1. 1Covalent labeling of protein components of the sodium channel with a photoactivable derivative of scorpion toxin.1980 · 202 citations
  2. 2Neurotoxins that Act on Voltage-Sensitive Sodium Channels in Excitable Membranes1980 · 1,247 citations
  3. 3Derivation of specific antibody‐producing tissue culture and tumor lines by cell fusion1976 · 1,927 citations
  4. 4A new method for labelling saxitoxin and its binding to non‐myelinated fibres of the rabbit vagus, lobster walking leg, and garfish olfactory nerves.1976 · 195 citations