Vector ConstructionThe dual-specificity MAPK phosphatase-3, ERK1/2 specific, mouse cDNA 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 mouse 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 (shown to reverse ERK1/2 activity)
Sanna B, Bueno OF, Dai Y-S, Wilkins BJ, Molkentin JD. Direct and indirect interactions between calcineurin-NFAT and MEK1-ERK1/2 signaling pathways regulate gene expression and cellular growth. Mol. Cell. Biol. 25:865-878. 2005.
Recombinant adenovirus was generated as described in Gomez-Foix, AM, et al., J. Biol. Chem. (1992)267:25129-25134.