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 Sc Endogenous 46, 54 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  Sc=S. cerevisiae
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

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 is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr183/Tyr185 of human SAPK/JNK.

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

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