Product Pathways - MAPK Signaling
c-Jun Fusion Protein #6093
Molecular Weight
37
Description
c-Jun Fusion Protein serves as a useful substrate for SAPK/JNK, which will phosphorylate it at Ser63 and Ser73 (1). It is expressed as a recombinant protein fusion of c-Jun residues 1-89 and glutathione S-transferase (1,2).
Western Blotting
Figure 1. Western blot analysis of c-Jun fusion protein phosphorylated by JNK1 kinase, using c-Jun (60A8) Rabbit mAb #9165 (upper panel) and Phospho-c-Jun (Ser63) (54B3) Rabbit mAb #2361 (lower panel).
Kinase Assay - Radiometric
Figure 2. c-Jun fusion protein was used as substrate to measure JNK1 kinases activity in a radiometric assay using the following reaction conditions: 25 mM Tris-HCl (pH7.5), 10 mM MgCl2, 5 mM β-glycerophosphate, 0.1 mM Na3VO4, 2 mM DTT, 50 μM ATP, Substrate: c-Jun fusion protein 400 ng/μL, and variable amounts of JNK1 kinases.
Directions for Use
c-Jun Fusion Protein at a concentration of 2 µg/20 µl reaction can be phosphorylated using 50 units of active SAPK in an in vitro kinase assay with 1X Kinase Buffer (#9802) and 200 µM ATP (#9804). After a 30-minute assay at 30°C, phosphorylation can be detected by Western blot with phospho-specific c-Jun antibodies (#9164 and #9261).
Background
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).
- Binetruy, B. et al. (1991) Nature 351, 122-127.
- Smeal, T. et al. (1991) Nature 354, 494-496.
- Derijard, B. et al. (1994) Cell 76, 1025-1037.
- Kyriakis, J.M. et al. (1994) Nature 369, 156-160.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 9164 Phospho-c-Jun (Ser73) Antibody
- 9261 Phospho-c-Jun (Ser63) II Antibody
- 9802 Kinase Buffer (10X)
- 9804 ATP (10 mM)
- 9810 SAPK/JNK Kinase Assay Kit (Nonradioactive)
- 9165 c-Jun (60A8) Rabbit mAb
- 7418 JNK1 Kinase
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
