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

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

Citations (1)

Simple Western™ analysis of lysates (0.1 mg/mL) from 293 cells treated with UV (50mJ, 0.5 hour recovery) using Phospho-SAPK/JNK (Thr183/Tyr185) Antibody #9251. The virtual lane view (left) shows two target bands (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Simple Western™ analysis of lysates (1 mg/mL) from 293 cells using SAPK/JNK Antibody #9252. The virtual lane view (left) shows two target bands (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from 293 or SK-N-MC cells treated with UV (40 mJ/cm2), using Phospho-SAPK/JNK (Thr183/Tyr185) Antibody (upper) or SAPK/JNK Antibody (lower).
Western blot analysis of extracts from 293 and SK-N-MC cells, untreated or UV-treated (40 J/m2), using Phospho-SAPK/JNK Antibody #9251 (upper) or SAPK/JNK Antibody (lower).
Western blot analysis of extracts from 293 cells, treated with or without UV light, using Phospho-SAPK/JNK (Thr183/Tyr185) (98F2) Rabbit mAb (upper) and SAPK/JNK (56G8) Rabbit mAb (lower).
Inquiry Info.# 9250

Kit Usage Information

Protocols

Product Description

The PhosphoPlus® SAPK/JNK (Thr183/Tyr185) Antibody Kit provides reagents and protocols for the rapid analysis of SAPK/JNK phosphorylation.

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 unphosphorylated SAPK/JNK. This antibody may slightly cross-react with phospho-ERK1/2 or -p38 phosphorylated at homologous residues. SAPK/JNK Antibody detects endogenous levels of total SAPK/JNK protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr183/Tyr185 of human SAPK/JNK or with a recombinant human JNK2 fusion protein. Antibodies are purified by protein A and peptide affinity chromatography.

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).

Pathways

Explore pathways related to this product.

Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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