c-Jun is a component of the transcription factor AP-1, which binds and activates transcription at TRE/AP-1 elements. The transcriptional activity of c-Jun is regulated by phosphorylation at Ser63 and Ser73 (1,2). Extracellular signals including growth factors, transforming oncoproteins and UV irradiation stimulate phosphorylation of c-Jun at Ser63/73 and activate c-Jun-dependent transcription. Mutation of Ser63/73 renders c-Jun nonresponsive to mitogenic and stress-induced signaling pathways. The MAP kinase homologue SAPK/JNK binds to the amino-terminal region of c-Jun and phosphorylates c-Jun at Ser63/73. In addition, the activity of SAPK/JNK is stimulated by the same signals that activate c-Jun (3,4).
Capture and Detection antibodies (100X stocks) and a HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are provided for performing 4 x 96 well ELISAs. Phospho-c-Jun (Ser63) Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added, followed by c-Jun Detection Antibody and HRP-conjugated secondary antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance at 450nm is proportional to the quantity of phospho-c-jun (Ser63) protein.
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