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Reisolation of immunoreactive radioiodinated antigens using glutaraldehyde-insolubilized antibody preparations
Radioiodination of antigens for use in radioimmunoassay can result in substantial losses of antigenic reactivity with the corresponding antibody and antisera preparations. We describe a method whereby antigens iodinated with the chloramine-T procedure are bound to and eluted from glutaraldehyde-insolubilized antibody. Unfractionated antisera, an ammonium sulfate precipitated fraction or the IgG fraction of antisera may be used as insolubilized immunoadsorbents. The method has been applied for the reisolation of a radioiodinated peptide, a low molecular weight protein and the fibronectin molecule. The total binding of 125I-antigens to antibody in radioimmunoassays can be increased from such low amounts before reisolation that the assay is not feasible, to 85% above background binding, after adsorption and elution from the insolubilized antibody preparations.
Radioiodination of antigens for use in radioimmunoassay can result in substantial losses of antigenic reactivity with the corresponding antibody and antisera preparations. We describe a method whereby antigens iodinated with the chloramine-T procedure are bound to and eluted from glutaraldehyde-insolubilized antibody. Unfractionated antisera, an ammonium sulfate precipitated fraction or the IgG fraction of antisera may be used as insolubilized immunoadsorbents. The method has been applied for the reisolation of a radioiodinated peptide, a low molecular weight protein and the fibronectin molecule. The total binding of 125I-antigens to antibody in radioimmunoassays can be increased from such low amounts before reisolation that the assay is not feasible, to 85% above background binding, after adsorption and elution from the insolubilized antibody preparations.