Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb #9255

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC F H M R Hm Endogenous 46 Phospho-JNK1. 54 Phospho-JNK2/3. Mouse IgG1

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb detects endogenous levels of p46 and p54 SAPK/JNK dually phosphorylated at Thr183 and Tyr185. This antibody does not recognize endogenous levels of phosphorylated p44/42 MAPK or p38 MAP kinase.

Source / Purification

Monoclonal antibody (isotype: mouse IgG1) is produced by immunizing mice with a synthetic phospho-peptide (KLH coupled) corresponding to residues surrounding Thr183/Tyr185 of human SAPK/JNK. The antibody is purified by affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, untreated or UV-treated (lanes 1 and 2), NIH/3T3 cells, untreated or UV-treated (lanes 3 and 4) and C6 cells, untreated or anisomycin-treated (lanes 5 and 6), using Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (green) or anisomycin-treated (blue), using Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells untreated (left) and anisomycin-treated (right) using Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb (green). Actin filaments have been labeled with DY554 phalloidin (red).


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.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

Products