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Molecular characterization of a membrane protein by a simple immunobinding procedure with monoclonal antibodies

A Sonnenberg, G J Kolvenbag, E J Al

    Journal of Immunological Methods
    |September 4, 1984
    PubMed

    A simple immunobinding procedure for the detection and molecular characterization of antigens is described. Antigen is adsorbed by immobilized antibodies, and this is followed by radiolabeling with iodine. Both adsorption and radioiodination are carried out in microtiter wells. After gel electrophoresis and autoradiography the apparent molecular weight of the radiolabeled antigen may be estimated. With this procedure we show that 2 monoclonal antibodies, directed against different determinants, both detect a glycoprotein with an apparent molecular weight of 170,000. By a 2-site sandwich immunoassay we demonstrate that these antibodies detect the same glycoprotein.

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