The PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit provides reagents and protocols for the rapid analysis of the phosphorylation status of p44 and p42 (Erk1 and Erk2).
p44/42 MAPK (Erk1/2) Negative Control Cell Extracts: Total cell extracts from Jurkat cells treated with U0126 (MEK1/2 Inhibitor) #9903 at 10 μM for 1 hour, to serve as a negative control. Supplied in SDS Sample Buffer.
p44/42 MAPK (Erk1/2) Positive Control Cell Extracts: Total cell extracts from Jurkat cells treated with TPA #4174 at 200 nM for 20 minutes, to serve as a positive control. Supplied in SDS Sample Buffer.
Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) Rabbit mAb #4370 detects endogenous levels of p42 and p44 MAP Kinase (Erk1 and Erk2) when dually phosphorylated at Thr202 and Tyr204 of Erk1 (Thr185 and Tyr187 of Erk2), and singly phosphorylated at Thr202. This antibody does not cross-react with the corresponding phosphorylated residues of either JNK/SAPK or p38 MAP kinases. p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 detects endogenous levels of total p44/42 MAP kinase (Erk1/Erk2) protein. The antibody does not cross-react with either JNK/SAPK or p38 MAP kinases.
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr202/Tyr204 of human p44 MAP kinase or a synthetic peptide corresponding to residues near the C-terminus of rat p44 MAP kinase.
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 research investigators consider it 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.
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