Phospho-p44/42 MAPK (Erk1/2) (

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Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb (Alexa Fluor^® 647 Conjugate) #4284

Applications	Reactivity	Sensitivity	Isotype
F	H M R Hm Mk Mi Dm Z B Pg	Endogenous	Rabbit IgG

Applications Key:  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  Mi=Mink  Dm=D. melanogaster  Z=Zebrafish  B=Bovine  Pg=Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

4284:

Flow

Specificity / Sensitivity

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb (Alexa Fluor^® 488 Conjugate) detects endogenous levels of p44 and p42 MAP Kinase (Erk1 and Erk2) when either dually phosphorylated at Thr202 and Tyr204 of Erk1 (Thr185 and Tyr187 of Erk2), or singly phosphorylated at Thr202. This antibody does not cross-react with the corresponding phosphorylated residues of either JNK/SAPK or p38 MAP kinases.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr202/Tyr204 of human p44 MAP kinase. 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.

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor^® 647 fluorescent dye and tested in-house for direct flow cytometric analysis of human cells. The unconjugated antibody #4370 reacts with human, mouse, rat. monkey, mink, pig, hamster, bovine, D. melanogaster and zebra fish phospho p44/p42 protein. CST expects that Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb (Alexa Fluor^® 647 Conjugate) will also recognize phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) 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 Pouysségur, 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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Companion Products

• 4344 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb (Alexa Fluor^® 488 Conjugate)
• 4370 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb
• 4377 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (197G2) Rabbit mAb
• 9106 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (E10) Mouse mAb
• 9101 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody
• 4695 p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb
• 4696 p44/42 MAPK (Erk1/2) (L34F12) Mouse mAb
• 9102 p44/42 MAPK (Erk1/2) Antibody
• 3452 Rabbit IgG Isotype Control (Alexa Fluor^® 647 Conjugate)
• 2985 Rabbit (DA1E) mAb IgG XP^® Isotype Control (Alexa Fluor^® 647 Conjugate)

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