Product Pathways - MAPK Signaling

Phospho-SAPK/JNK (Thr183/Tyr185) Antibody #9251

Applications	Reactivity	MW (kDa)	Source
W IP IHC-P IF-IC F	H M R Mk Hm B Dr (X)	46 (Phospho-JNK1). 54 (Phospho-JNK2/3).	Rabbit

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 Hm=Hamster B=Bovine Dr=Drosophila X=Xenopus
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-SAPK/JNK (Thr183/Tyr185) Antibody detects endogenous levels of p46 and p54 SAPK/JNK dually phosphorylated at threonine 183 and tyrosine 185. This antibody does not recognize endogenous levels of phosphorylated p44/42 MAPK or p38 MAP kinase.

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.

Western Blotting

Western blot analysis of extracts from 293 or SK-N-MC cells treated with UV (40 J/m2), using Phospho-SAPK/JNK (Thr183/Tyr185) Antibody (upper) or control SAPK/JNK (Thr183/Tyr185) antibody (lower).

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 (center) and Phospho-p38 MAPK Antibody #9211 (right).

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using Phospho-SAPK/JNK (Thr183/Tyr185) Antibody.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Phospho-SAPK/JNK (Thr183/Tyr185) Antibody.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or calf intestinal phosphatase (CIP)-treated (right), using Phospho-SAPK/JNK (Thr183/Tyr185) Antibody.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-SAPK/JNK (Thr183/Tyr185) Antibody.

IHC-FL (floating)

Immunofluorescent analysis of COS-1 cells, untreated (right) or EGF-treated (100 ng/ml for 15 minutes, left), using Phospho-SAPK/JNK (Thr183/Tyr185) Antibody (green) and Texas Red-X-conjugated phalloidin (red) to detect actin. EGF stimulation caused activation and nuclear localization of JNK and rearrangement of the actin cytoskeleton into numerous ruffles at the periphery of the cells. (Provided by Harry Mellor, Ph.D., University of Bristol, UK.)

Flow Cytometry

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

IF-IC

Confocal immunofluorescent images of C6 cells, untreated (left) or anisomycin treated (right) labeled with Phospho-SAPK/JNK (Thr183/Tyr185) Antibody (green) and beta-Tubulin Antibody #2146 (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

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).

Application References

Kujime, K. et al. (2000) p38 mitogen-activated protein kinase and c-jun-NH2-terminal kinase regulate RANTES production by influenza virus-infected human bronchial epithelial cells. J. Immunol. 164, 3222-3228. This article references the use of Phospho-SAPK/JNK (Thr183/Tyr185) Antibody in the following applications: Western Blotting
Xiao, G. G. et al. (2003) Use of proteomics to demonstrate a hierarchical oxidative stress response to diesel exhaust particle chemicals in a macrophage cell line. J. Biol. Chem. 278, 50781-50790. This article references the use of Phospho-SAPK/JNK (Thr183/Tyr185) Antibody in the following applications: Western Blotting
Shukla, A. et al. (2001) Silica-induced activation of c-Jun-NH2-terminal amino kinases, protracted expression of the activator protein-1 proto-oncogene, fra-1, and S-phase alterations are mediated via oxidative stress. Cancer Res. 61, 1791-1795. This article references the use of Phospho-SAPK/JNK (Thr183/Tyr185) Antibody in the following applications: Western Blotting
Weiss, L. et al. (2000) Regulation of c-Jun NH(2)-terminal kinase (Jnk) gene expression during T cell activation. J. Exp. Med. 191, 139-146. This article references the use of Phospho-SAPK/JNK (Thr183/Tyr185) Antibody in the following applications: Flow Cytometry
Rochat-Steiner, V. et al. (2000) FIST/HIPK3: A Fas/fadd-interacting serine/threonine kinase that induces fadd phosphorylation and inhibits fas-mediated jun nh(2)-terminal kinase activation. J. Exp. Med. 192, 1165-1174. This article references the use of Phospho-SAPK/JNK (Thr183/Tyr185) Antibody in the following applications: Western Blotting

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

9250 PhosphoPlus® SAPK/JNK (Thr183/Tyr185) Antibody Kit
9252 SAPK/JNK Antibody
9253 SAPK/JNK Control Cell Extracts
9255 Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb
9810 SAPK/JNK Kinase Assay Kit (Nonradioactive)
9912 Phospho-SAPK/JNK Pathway Sampler Kit
7074 Anti-rabbit 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
1215 Phospho-SAPK/JNK (Thr183/Tyr185) Blocking Peptide