Product Pathways - MAPK Signaling
PhosphoPlus® SAPK/JNK (Thr183/Tyr185) Antibody Kit #9250
|9250S||1 Kit (20 western blots)||---||In Stock||---|
|9250||carrier free and custom formulation / quantity||email request|
|Kit Includes||Quantity||Applications||Reactivity||Mol. Wt.||Isotype|
|Phospho-SAPK/JNK (Thr183/Tyr185) Antibody #9251||200 µl||W, IP||H, M, R, Hm, Mk, Dm, B, Sc||46, 54 kDa||Rabbit|
|SAPK/JNK (56G8) Rabbit mAb #9258||200 µl||W||H, M, R, Hm, Mk, Mi||46, 54 kDa||Rabbit|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||W||All||Goat|
|Anti-biotin, HRP-linked Antibody #7075||250 µl||Goat|
|20X LumiGLO® Reagent and 20X Peroxide #7003||10 ml||W|
|Biotinylated Protein Ladder Detection Pack #7727||250 µl|
|SAPK/JNK Control Cell Extracts #9253||150 µl|
Applications Key: W=Western Blotting, IP=Immunoprecipitation
Reactivity Key: H=Human, M=Mouse, R=Rat, Hm=Hamster, Mk=Monkey, Dm=D. melanogaster, B=Bovine, Sc=S. cerevisiae, Mi=Mink, All=All Species Expected
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from 293 or SK-N-MC cells treated with UV (40 mJ/cm2), using Phospho-SAPK/JNK (Thr183/Tyr185) Antibody (upper) or control SAPK/JNK (Thr183/Tyr185) antibody (lower).
The PhosphoPlus® SAPK/JNK (Thr183/Tyr185) Antibody Kit provides reagents and protocols for the rapid analysis of SAPK/JNK phosphorylation.
Specificity / Sensitivity
Phospho-SAPK/JNK (Thr183/Tyr185) Antibody detects the dually phosphorylated isoforms of all three SAPKs/JNKs. Western analysis shows that this antibody does not cross-react with endogenous levels of the corresponding phosphorylated forms of p44/42 MAP Kinase or p38 MAP Kinase. However, because of the close homology between the active sites of SAPK/JNK and p44/42 MAP Kinase, larger amounts of phospho-p44/42 MAP Kinase may be detectable.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr183/Tyr185 of human SAPK/JNK. Antibodies are purified by protein A and peptide affinity chromatography.
Monoclonal antibody is produced by immunizing animals with a human JNK2/MBP fusion protein.
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.
- Watanabe, H. et al. (2001) J. Biol. Chem. 276, 14466-14473. Applications: Western Blotting.
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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
U.S. Patent No. 5,675,063.
PhosphoPlus® is a trademark of Cell Signaling Technology, Inc.
LumiGLO® is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.