Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb (Alexa Fluor® 647 Conjugate) #9257

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

Applications Key:  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) mAb (Alexa Fluor® 647 Conjugate) 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 around Thr183/Tyr185 of human SAPK/JNK. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions with an F/P ratio of 2-6. This antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-6. The Alexa Fluor® 647 dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugates of the Alexa Fluor® 647 dye produce bright far-red-fluorescence emission, with a peak at 665 nm.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of THP1 cells, untreated (blue) or anisomycin-treated (green), using Phospho-SAPK/JNK (T183/Y185) (G9) Mouse mAb (Alexa Fluor®647 Conjugate) (#9257) compared to a nonspecific control antibody (red).

Description

Cell Signaling Technology Antibody conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct Flow Cytometric analysis of human and mouse cells. The unconjugated antibody #9255 reacts with, among others, human, mouse, rat and hamster phospho-SAPK/JNK (Thr183/Tyr185). CST expects that Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb (Alexa Fluor® 647 Conjugate) will also recognize phospho-SAPK/JNK (Thr183/Tyr185) in these species.

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

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

The Alexa Fluor® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc., for research use only, except for use in combination with DNA microarrays. The Alexa Fluor® dyes (except for Alexa Fluor® 430 Dye) are covered by pending and issued patents.Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

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

Products