We describe generation of stable, fluorescence-barcoded cell lines suitable for multiplex screening of antibody to membrane proteins. The utility of this cell-based system, capable of a 256-plex cell panel, is demonstrated by flow cytometry deconvolution of barcoded cell panels expressing influenza A hemagglutinin trimers, or native human CCR2 or CCR5 multi-spanning proteins and their epitope-defining mutants. This platform may prove useful for characterizing immunity and discovering antibodies to membrane-associated proteins.