Cell Signaling Technology

Product Pathways - MAPK Signaling

Immobilized Phospho-p44/42 MAPK (Thr202/Tyr204) Mouse mAb #9109

Applications Reactivity MW (kDa) Source Isotype
IP H M R 42, 44 Mouse IgG2a

Applications Key:  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Immobilized Phospho-p44/42 MAPK (Thr202/Tyr204) Mouse mAb binds only p44/42 MAP kinase activated by phosphorylation at Thr202/Tyr204. It is useful for immunoprecipitation kinase assays. The antibody does not significantly cross-react with SAPK/JNK or p38 MAP kinase phosphorylated at the corresponding residue or with nonphosphorylated MAPK.

Source / Purification

Monoclonal antibody is produced by immunizing mice with a synthetic phospho-peptide (KLH-coupled) corresponding to residues around Thr202/Tyr204 of human p44 MAPK. Antibodies are immobilized by conjugation of carbohydrates to cross-linked agarose hydrazide beads.

IP

IP

MAP kinase activity was analyzed in anisomycin-treated or untreated NIH/3T3 cell extracts. Cell extracts were immunoprecipitated with Immobilized Phospho-p44/42 MAP Kinase (Thr202/Tyr204) Mouse mAb. In vitro kinase assays were performed using Elk-1 as a substrate. Phosphorylation of Elk-1 at Ser383 was visualized by immunoblotting with Phospho-Elk-1 (Ser383) Antibody #9181.

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), a MAP kinase kinase (MAPKK), and a MAP kinase. While multiple ERK1/2 MAP3Ks have been identified, including the Raf family, Mos, and Tpl2/Cot, MEK1 and MEK2 are the primary MAPKKs in this pathway (5,6). MEK1 and MEK2 activate ERK1/p44 and ERK2/p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of ERK1/2 have been identified, including p90RSK (7) and the transcription factor Elk-1 (8,9). ERK1/2 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

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

Product Pathways

Drug Discovery Tools

Featured Technologies

Protein Classes