Product Pathways - MAPK Signaling
SAPK/JNK Control Cell Extracts #9253
Protocols
- 9253:
- Western Blotting
Description
Nonphosphorylated SAPK/JNK Control Cell Extracts: Total cell extracts from 293 cells, prepared without treatment, serve as a negative control. Supplied in SDS Sample Buffer.Phosphorylated SAPK/JNK Control Cell Extracts: Total cell extracts from 293 cells, treated with UV light, serve as a positive control. Supplied in SDS Sample Buffer.
Applications
As controls, we recommend using 15 µl of phosphorylated and nonphosphorylated SAPK/JNK control extracts. Boil 2 minutes prior to use.
Background
The stress-activated protein kinase/Jun-amino-terminal kinase SAPK/JNK is potently and preferentially activated by a variety of environmental stresses including UV and gamma radiation, ceramides, inflammatory cytokines, and in some instances, growth factors and GPCR agonists (1-6). As with the other MAPKs, the core signaling unit is composed of a MAPKKK, typically MEKK1-MEKK4, or by one of the mixed lineage kinases (MLKs), which phosphorylate and activate MKK4/7. Upon activation, MKKs phosphorylate and activate the SAPK/JNK kinase (2). Stress signals are delivered to this cascade by small GTPases of the Rho family (Rac, Rho, cdc42) (3). Both Rac1 and cdc42 mediate the stimulation of MEKKs and MLKs (3). Alternatively, MKK4/7 can be activated in a GTPase-independent mechanism via stimulation of a germinal center kinase (GCK) family member (4). There are three SAPK/JNK genes each of which undergoes alternative splicing, resulting in numerous isoforms (3). SAPK/JNK, when active as a dimer, can translocate to the nucleus and regulate transcription through its effects on c-Jun, ATF-2, and other transcription factors (3,5).
- Davis, R.J. (1999) Biochem Soc Symp 64, 1-12.
- Ichijo, H. (1999) Oncogene 18, 6087-93.
- Kyriakis, J.M. and Avruch, J. (2001) Physiol Rev 81, 807-69.
- Kyriakis, J.M. (1999) J Biol Chem 274, 5259-62.
- Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
- Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem Sci 23, 481-5.
Application References
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Companion Products
- 9250 PhosphoPlus® SAPK/JNK (Thr183/Tyr185) Antibody Kit
- 9251 Phospho-SAPK/JNK (Thr183/Tyr185) Antibody
- 9252 SAPK/JNK Antibody
- 9255 Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 4668 Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb
- 9258 SAPK/JNK (56G8) Rabbit mAb
- 4671 Phospho-SAPK/JNK (Thr183/Tyr185) (98F2) Rabbit mAb
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
For Research Use Only. Not For Use In Diagnostic Procedures.