Vector ConstructionDominant negative AGF mutation in human p38alpha with a 5’ flag epitope was cloned into the multiple cloning site of pAC-CMV-pLpA. The resulting plasmid was cotransfected into HEK293 cells with plasmid pJM17 which contains the Ad5 genome. Homologous recombination between the two plasmids resulted in the replacement of the Ad5 early region 1 with the human cDNA expression cassette, generating a replication deficient recombinant virus.
Supplied AsCrude HEK293 cell lysate.
StorageStore at -80°C. Multiple freeze/thaw cycles not recommended.
QC TestingWestern Blot (to block endogenous p38 phos)
Kaiser RA, Bueno OF, Lips DJ, Doevendans PA, Kimball TF, Molkentin JD . Targeted Inhibition of p38 MAPK Antagonizes Cardiac Injury and Cell Death Following Ischemia-Reperfusion in vivo. J. Biol. Chem. 279:15524-15530. 2004. Recombinant adenovirus was generated as described in Gomez-Foix, AM, et al., J. Biol. Chem. (1992)267:25129-25134.