Cell Signaling Technology

Product Pathways - MAPK Signaling

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

Kit Includes Quantity Applications Reactivity MW (kDa) Source
Phospho-SAPK/JNK (Thr183/Tyr185) Antibody # 9251 200 microliters W IP IHC-P IF-IC F H M R Mk Hm B Dr (X) 46 (Phospho-JNK1). 54 (Phospho-JNK2/3). Rabbit
SAPK/JNK (56G8) Rabbit mAb # 9258 200 microliters W H M R Mk Mi Hm 46, 54 Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074 100 microliters Goat
Anti-biotin, HRP-linked Antibody # 7075 200 microliters Goat
20X LumiGLO® Reagent and 20X Peroxide # 7003 10 milliliters
Biotinylated Protein Ladder Detection Pack # 7727 200 microliters
SAPK/JNK Control Cell Extracts # 9253 60 microliters

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Mi=Mink  Hm=Hamster  B=Bovine  Dr=Drosophila  X=Xenopus

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).

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH coupled) 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 rabbits 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, by growth factors and GPCR agonists (1-6). As with the other MAPKs, the core signaling unit is composed of a MAPKKK, typically MEKK1-4, or by one of the mixed lineage kinases (MLKs), which phosphorylate and activate MKK4-7, which then 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 by a pathway independent of small GTPases via stimulation of a member of the germinal center kinase (GCK) family (4). There are three SAPK/JNK genes with further diversification resulting from alternative splicing (3). Active SAPK/JNK dimers can translocate to the nucleus to 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, 6087-6093.
  3. Kyriakis, J.M. and Avruch, J. (2001) Physiol. Rev. 81, 807-869.
  4. Kyriakis, J.M. (1999) J. Biol. Chem. 274, 5259-5262.
  5. Leppa, S. and Bohmann, D. (1999) Oncogene 18, 6158-6162.
  6. Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem. Sci. 23, 481-485.

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