C-Jun is a component of the transcription factor AP-1 that 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 N-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). Nonphosphorylated c-Jun (Ser63) II Cell Extracts: Total cell extracts from NIH/3T3 cells, prepared without treatment, serve as a negative control.
Kit Compontents:
C5815-08A Non-phosphorylated c-Jun control extract. 1x150ul Supplied as a liquid in 62.5 mM Tris-HCl, pH 6.8, 2% SDS, 10% glycerol, 50 mM DTT, 0.01% phenol red.
C5815-0B Phosphorylated c-Jun control extract.1x150ul Supplied as a liquid in 62.5 mM Tris-HCl, pH 6.8, 2% SDS, 10% glycerol, 50 mM DTT, 0.01% bromophenol blue
Applications:
Suitable for use in Western Blot. Other applications not tested.
Recommended Dilution:
Western Blot: As controls, use15ul of C5815-08A non-phosphorylated and C5815-08B phosphorylated c-Jun control extracts.
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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