Evaluation of an enzyme immunoassay kit for estrogen receptor measurements.
Using an enzyme immunoassay (EIA) procedure, we evaluated the ability of the glass beads in the ER-EIA kit of Abbott Laboratories, which were coated with monoclonal anti-estrogen receptor (ER) antibodies, to bind hormone-free and hormone-filled Type I ER in cytosols, buffer washes, and 0.4 mol/L KCI extracts of ultracentrifugal pellets of breast cancer tissue homogenates. The unmodified ER-EIA technique yields higher values than does the dextran-coated charcoal (DCC) method for Type I ER. However, the antibody-coated beads fail to bind hormone-free ER and react with only a certain proportion of ER-[3H]estradiol complexes. The antigen saturation limit of the beads could not be determined because the quantity of antigen bound by the beads was disproportionate and unrelated to the total amount of the antigen available in the cytosols. In buffer extracts of tissue pellets that were ER-negative by the DCC assay, the EIA method detected high quantities of ER. We recommend checking the ability of the monoclonal antibodies to recognize proteins other than Type I ER in the extra-nuclear and nuclear compartments of target cells before using them for immunohistochemical detection of ER.
D000911:Antibodies, Monoclonal / Q000276:immunology; D001943:Breast Neoplasms / Q000032:analysis; D002606:Charcoal; D003911:Dextrans; D006801:Humans; D007124:Immunoenzyme Techniques; D008982:Molybdenum / Q000494:pharmacology; D009994:Osmolar Concentration; D011933:Reagent Kits, Diagnostic; D011960:Receptors, Estrogen / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D003911:Dextrans; D011933:Reagent Kits, Diagnostic; D011960:Receptors, Estrogen; D002606:Charcoal; D008982:Molybdenum; C024687:sodium molybdate(VI)