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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).

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Western blot analysis of extracts from HeLa, NIH/3T3, PC12 and COS cells, using SAPK/JNK (56G8) Rabbit mAb.

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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.

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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.

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-SAPK/JNK (Thr183/Tyr185) Antibody 9251 200 µl
Western Blotting Immunoprecipitation
H M R Hm Mk Dm B Sc 46, 54 Rabbit 
SAPK/JNK (56G8) Rabbit mAb 9258 200 µl
Western Blotting
H M R Hm Mk Mi 46, 54 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
Western Blotting
All Goat 
Anti-biotin, HRP-linked Antibody 7075 250 µl
Goat 
20X LumiGLO® Reagent and 20X Peroxide 7003 10 ml
Western Blotting
 
Biotinylated Protein Ladder Detection Pack 7727 250 µl
 
SAPK/JNK Control Cell Extracts 9253 150 µl
 

Product Description

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).


1.  Ichijo, H. (1999) Oncogene 18, 6087-93.

2.  Kyriakis, J.M. and Avruch, J. (2001) Physiol Rev 81, 807-69.

3.  Kyriakis, J.M. (1999) J Biol Chem 274, 5259-62.

4.  Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem. Sci. 23, 481-485.

5.  Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.

6.  Davis, R.J. (1999) Biochem Soc Symp 64, 1-12.


Entrez-Gene Id 5599
Swiss-Prot Acc. P45983


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
PhosphoPlus® is a trademark of Cell Signaling Technology, Inc.
LumiGLO® is a registered trademark of Kirkegaard & Perry Laboratories.
U.S. Patent No. 5,675,063.