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Polyclonal antibodies to Akt recognize all three known isoforms of Akt. COS-7 cells cultured in 6-cm-diameter plates were transiently transfected with 3 μg of plasmid-encoded FLAG-tagged Akt1, Akt2, or RAC-PKγ. Lysates prepared from COS-7 cells expressing each isoform of Akt were subjected to immunoprecipitation with polyclonal antibodies to Akt. The immunoprecipitates (lanes 1 to 3) or the total cell lysates (lanes 4 to 6) were subjected to immunoblot analysis with antibodies to FLAG.
