A radioimmunoassay with monoclonal antibodies for the detection of antigenic cell-free Fc receptor.
Making use of 125I-labelled monoclonal rat antibodies against mouse Fc receptor we have developed a solid-phase radioimmunoassay for cell-free antigenic mouse Fc receptor (FcR). An 80% acetone solution was used for fixation of relatively large amounts of soluble proteins on PVC microtiter plates. As a result of this treatment FcR practically lost its activity to bind the Fc portion of an IgG molecule but retained its antigenicity, the binding of specific anti-FcR (aFcR) antibody being increased following acetone fixation. Concentrations of cell-free FcR in NP-40 extracts of FcR-expressing cells were calculated from the linear part of a standard curve and expressed in units of antigenic activity, 1 unit being the amount of antigenic FcR capable of binding 1 microgram of 125I-aFcR. The method may be used for detecting cell-free FcR as a minor constituent in a mixture of proteins.
D000096:Acetone / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001666:Binding Sites, Antibody; D002457:Cell Extracts / Q000378:metabolism; D002460:Cell Line; D000939:Epitopes / Q000032:analysis*; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011863:Radioimmunoassay / Q000379:methods* / Q000592:standards; D011961:Receptors, Fc / Q000032:analysis* / Q000187:drug effects / Q000276:immunology
D000911:Antibodies, Monoclonal; D002457:Cell Extracts; D000939:Epitopes; D011961:Receptors, Fc; D000096:Acetone