Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) Mouse mAb (Alexa Fluor® 647 Conjugate) #4375

Applications Reactivity Sensitivity Isotype
F H M Endogenous Mouse IgG1kappa

Applications Key:  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) Mouse mAb (Alexa Fluor® 647 conjugate) detects endogenous levels of p44 and p42 MAP kinase (Erk1 and Erk2) dually phosphorylated at threonine 202 and tyrosine 204. This antibody does not cross-react with the corresponding phosphorylated residues of either SAPK/JNK or p38 MAP kinase.

Source / Purification

Monoclonal antibody (isotype: mouse IgG1) is produced by immunizing mice with a synthetic phospho-peptide (KLH-coupled) corresponding to residues around Thr202/Tyr204 of human p44 MAP kinase. Antibody was conjugated with Alexa Fluor® 647 under optimum conditions with a F/P ratio of 2-6.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, UO126-treated (blue) or PMA-treated (green), using Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) Mouse mAb (Alexa Fluor® 647 conjugate) 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 #9106 reacts with, among others, human, mouse, rat, hamster and Xenopus p44/42 MAPK. CST expects that Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) Mouse mAb (Alexa Fluor® 647 conjugate) will also recognize p44/42 MAPK in these species.

Background

Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs such as cell proliferation, differentiation, motility, and death. The p44/42 MAPK (ERK1/2) signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens, growth factors, and cytokines (1-3) and is an important target in the diagnosis and treatment of cancer (4). Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK or MAP3K), a MAP kinase kinase (MAPKK or MAP2K), and a MAP kinase (MAPK). Multiple p44/42 MAP3Ks have been identified, including members of the Raf family as well as Mos and Tpl2/Cot. MEK1 and MEK2 are the primary MAPKKs in this pathway (5,6). MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of p44/42 have been identified, including p90RSK (7) and the transcription factor Elk-1 (8,9). p44/42 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs (10), along with MEK inhibitors such as U0126 and PD98059.

  1. Roux, P.P. and Blenis, J. (2004) Microbiol Mol Biol Rev 68, 320-44.
  2. Baccarini, M. (2005) FEBS Lett 579, 3271-7.
  3. Meloche, S. and Pouyssegur, J. (2007) Oncogene 26, 3227-39.
  4. Roberts, P.J. and Der, C.J. (2007) Oncogene 26, 3291-310.
  5. Rubinfeld, H. and Seger, R. (2005) Mol Biotechnol 31, 151-74.
  6. Murphy, L.O. and Blenis, J. (2006) Trends Biochem Sci 31, 268-75.
  7. Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
  8. Marais, R. et al. (1993) Cell 73, 381-93.
  9. Kortenjann, M. et al. (1994) Mol Cell Biol 14, 4815-24.
  10. Owens, D.M. and Keyse, S.M. (2007) Oncogene 26, 3203-13.

Application References

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Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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