Cell Signaling Technology

Product Pathways - MAPK Signaling

PhosphoPlus® SAPK/JNK (Thr183/Tyr185) Antibody Kit #9250

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

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  Mi=Mink  Dm=D. melanogaster  X=Xenopus  B=Bovine  Sc=S. cerevisiae
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

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.

Western Blotting

Western Blotting

Phospho SAPK/JNK (Thr183/Tyr185) Antibody recognizes p54/p46 SAPK/JNK but not Erk1/2 or p38 MAPK. Western blot analysis of C6 cell extracts with and without anisomycin treatment, using Phospho-ERK1/2 Antibody #9101 (left), Phospho-SAPK/JNK Antibody #9251 (center) and Phospho-p38 MAPK Antibody #9211 (right).

Description

The PhosphoPlus® SAPK/JNK (Thr183/Tyr185) Antibody Kit provides reagents and protocols for the rapid analysis of SAPK/JNK phosphorylation.

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.

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).

  1. Davis, R.J. (1999) Biochem Soc Symp 64, 1-12.
  2. Ichijo, H. (1999) Oncogene 18, 6087-93.
  3. Kyriakis, J.M. and Avruch, J. (2001) Physiol Rev 81, 807-69.
  4. Kyriakis, J.M. (1999) J Biol Chem 274, 5259-62.
  5. Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
  6. Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem Sci 23, 481-5.

Application References

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Protocols

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For Research Use Only. Not For Use In Diagnostic Procedures.

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