Product Pathways - MAPK Signaling
Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb #9255
PhosphoSitePlus® protein, site, and accession data: JNK1
| 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 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 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
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).
- Davis, R.J. (1999) Biochem Soc Symp 64, 1-12.
- Ichijo, H. (1999) Oncogene 18, 6087-93.
- Kyriakis, J.M. and Avruch, J. (2001) Physiol Rev 81, 807-69.
- Kyriakis, J.M. (1999) J Biol Chem 274, 5259-62.
- Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
- Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem Sci 23, 481-5.
Application References
- Akagi, T. et al. (2000) v-Crk activates the phosphoinositide 3-kinase/AKT pathway in transformation. Proc.Natl. Acad.Sci. USA 97, 7290-7295. Applications: Western Blotting
- Song, J. S. et al. (1999) Tyrosine phosphorylation of Vav stimulates IL-6 production in mast cells by a Rac/c-Jun N-terminal kinase-dependent C208pathway. Journal of Immunology 163, 802-810. Applications: Western Blotting
- Zhang, S. and Kaplan, M.H. (2000) The p38 mitogen-activated protein kinaseis required for IL-12-induced IFN-gamma expression. Journal of Immunology 165, 1374-1380. Applications: Western Blotting
- Xing, J. et al. (1998) Nerve growth factor activates extracellular signal-regulated kinase and p38 mitogen-activated protein kinase pathways to stimulate CREB serine 133 phosphorylation. Molecular and Cellular Biology 18, 1946-1955. Applications: Western Blotting
- Saporito, M. S. et al. (2000) MPTP activates c-Jun NH(2)-terminal kinase (JNK) and its upstream regulatory kinase MKK4 in nigrostriatal neurons in vivo. J. Neurochem. 75, 1200-1208. Applications: Western Blotting
- Turner, N.A. et al. (2009) Am J Physiol Heart Circ Physiol 297, H1117-27. Applications: Western Blotting
- McPherson, V.A. et al. (2009) J Immunol 183, 4940-7. Applications: Western Blotting
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Companion Products
- 4668 Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb
- 9251 Phospho-SAPK/JNK (Thr183/Tyr185) Antibody
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- 9253 SAPK/JNK Control Cell Extracts
- 9258 SAPK/JNK (56G8) Rabbit mAb
- 7072 Phototope®-HRP Western Blot Detection System, Anti-mouse IgG, HRP-linked Antibody
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 9250 PhosphoPlus® SAPK/JNK (Thr183/Tyr185) Antibody Kit
- 9912 Phospho-SAPK/JNK Pathway Antibody Sampler Kit
- 5755 Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb (PE Conjugate)
For Research Use Only. Not For Use In Diagnostic Procedures.