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
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.
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Application References
- Dashti, S.R. et al. (2001) J Biol Chem 276, 8059-63. This article references the use of Immobilized Phospho-p44/42 MAPK (Thr202/Tyr204) Mouse mAb in the following applications: IP
- Ge, N.L. and Rudikoff, S. (2000) Blood 96, 2856-61. This article references the use of Immobilized Phospho-p44/42 MAPK (Thr202/Tyr204) Mouse mAb in the following applications: IP
- Chen, Y.J. et al. (2007) J Biol Chem 282, 27215-28. This article references the use of Immobilized Phospho-p44/42 MAPK (Thr202/Tyr204) Mouse mAb in the following applications: IP
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Companion Products
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