Determination of cell-bound Fc receptors using a cell adherence test (CAT).
The determination of membrane-bound Fc receptors as rosette forming cells with antibody-coated erythrocytes (EA-RFC) is a tedious and time consuming method. Here, we describe a modification of the assay delineated by Eisenbarth et al. (J. Immunol. Methods (1980) 39, 387) for the detection of Fc receptors on cell surfaces. V-shaped microtiter plates were coated with an antigen (e.g., fluorescein isothiocyanate (FITC) coupled to bovine serum albumin (BSA] and subsequently with monoclonal anti-FITC antibodies of different isotypes. Cells to be tested were added, incubated for a short time or centrifuged immediately. Cells expressing FcR, became bound to the flanks of the V-shaped wells, whereas cells lacking the receptor were pelleted to the tip of the well. This cell adherence test (CAT) is independent of the agglutinating abilities of the antibodies, and since it is very fast and highly reproducible, large numbers of different cells can be screened for FcR expression. Anti-receptor antibodies can also be tested for their binding to FcR.
Lemke
H
H
Biochemisches Institut, Medizinischen Fakultät, Christian-Albrechts-Universität zu Kiel, F.R.G.
Huss
G
G
Bonmann
J
J
Hansen
H
H
D000818:Animals; D002448:Cell Adhesion; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D011961:Receptors, Fc / Q000032:analysis* / Q000276:immunology; D012397:Rosette Formation; D013861:Thiocyanates
D005452:Fluoresceins; D011961:Receptors, Fc; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate