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A chimera containing the CD28 cytoplasmic domain localizes to lipid rafts and is tyrosine-phosphorylated. Lysates of unstimulated Jurkat T cells that stably express a Lck-CD28-FLAG chimera (described in Materials and Methods) were fractionated on a sucrose gradient. Anti-FLAG antibody was used to immunoprecipitate the chimera, which was then analyzed by immunoblotting with antibodies to phosphotyrosine and the FLAG epitope.
