Quantitative Determination Of Cell Surface Antigens By Indirect Immunofluorescence Assay Using Qifikit Calibrator Beads
In this procedure, the cell specimen is labeled with a saturating concentration of either a primary mouse monoclonal antibody (MAb) directed against the antigen of interest or an irrelevant mouse monoclonal antibody (control). Saturation conditions are determined through titration experiments for each MAb investigated, using a fluorescein-conjugated anti-mouse secondary antibody. The primary antibody may be of any mouse IgG isotype. Under these conditions, the number of bound primary antibody molecules corresponds to the number of antigenic sites present on the cell surface.
Contributed by Jean-Luc D'hautcourt
Current Protocols in Cytometry (2002) 6.12.1-6.12.22 Copyright © 2002 by John Wiley & Sons, Inc.
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