Prior to animal immunization, plasmids and adenoviruses encoding various influenza viral genes were tested for their expression in 293T and A549 cells. Plasmids encoding HA protein of A/Thailand/1/KAN-1/2004, NP proteins of A/PR/8/34 and A/Thailand/1/KAN-1/2004, and M2 protein of A/Thailand/1/ KAN-1/2004 were transferred into 293T cells using the calcium phosphate-mediated ProFectionH Mammalian Transfection system (Promega, Madison, WI). Adenoviruses encoding HA (KAN-1), NP (KAN-1) and M2 (KAN-1) were transfected into A549 cells for 48 hours followed by a change of media. Cell lysates were collected 48 hours post-transfection and subjected to Western blot analysis by mouse monoclonal antibodies against HA (KAN-1), NP (KAN-1), NP (PR/8), and by ferret anti-serum raised against M2 (KAN-1). Specific bands of the predicted size of proteins were detected by comparison to a known vector control.