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Fig. 4. Detection of cell surface FcRn. The cell surface of FO-1 cells (lane 3), FO-1ß2m cells (lane 5), FO-1 cells expressing Myc-hFcRn (lanes 1,4) and FO-1ß2m cells expressing Myc-hFcRn (lanes 2,6) were biotinylated on ice with a membrane-impermeable reagent. Cells were lysed and an aliquot of the lysates directly analyzed by immunoblotting (lysate; lanes 1,2) to detect the presence of Myc-hFcRn ({alpha}-Myc). The remaining lysate was precipitated with anti-Myc antibodies ({alpha}-Myc IP) and biotinylated Myc-hFcRn present in the precipitates detected by blotting with streptavidin-HRP (SA-HRP, lanes 3-6, top panel). Alternatively, biotinylated proteins were first precipitated with streptavidin-agarose (SA-P) and precipitates immunoblotted to detect Myc-hFcRn ({alpha}-Myc, lower panel). hc, heavy chain of the antibody used for immunoprecipitation. The data shown is representative of at least three independent experiments, each carried out using two different cell clones.