Product Pathways - MAPK Signaling

Growth/Proliferation Marker: SignalStain® Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Detection Kit #8110

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Kit Includes	Cap Color
Peroxidase Quench	Orange
Blocking Solution	Blue
Prediluted Phospho-p44/42 MAPK (Thr202/Tyr204) (20G11) Rabbit Monoclonal Antibody #4376	Purple
Prediluted Negative Control	Brown
Biotinylated Secondary Antibody	Green
Reagent A	Gray
Reagent B	Gray
NovaRED Substrate 1 (TM)	Red
NovaRED Substrate 2 (TM)	Red
NovaRED Substrate 3 (TM)	Red
NovaRED Substrate 4 (TM)	Red
Phospho-p44/42 MAPK (Thr202/Tyr204) Blocking Peptide # 1150	White

NovaRED™ is a trademark of Vector Laboratories.

Description

CST's Growth/Proliferation Marker: SignalStain® Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Detection Kit is a "ready to use" system designed to detect the activation of MAP kinase in human tissue and cell preps by immunohistochemistry. The kit utilizes the ABC immunoperoxidase method to detect endogenous levels of phosphorylated p44/42 MAP kinase. Prediluted Phospho-p44/42MAPK (Thr202/Tyr204) (20G11) Rabbit mAb #4376 is bound by a biotinylated secondary antibody. Avidin DH and biotinylated horseradish peroxidase are complexed by mixing in defined amounts prior to use, and the mixture subsequently binds the secondary antibody. The macromolecular complex is localized by incubation with NovaRED™ enzyme substrate.The prediluted primary antibody, along with the ABC system, allows the user consistently to examine phosphorylated-p44/42 MAP Kinase localization and offers the highest sensitivity with the lowest background.

Species Cross-Reactivity

H (M) (R) (Mk) (Z)

Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Z=Zebra Fish
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Growth/Proliferation Marker: SignalStain® Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Detection Kit recognizes endogenous levels of p42 and p44 MAP kinase (Erk1 and Erk2) only when phosphorylated at Thr202 and Tyr204 of Erk1 (Thr183 and Tyr185 of Erk2). The antibody does not cross-react with the corresponding phosphorylated residues of either SAPK/JNK or p38 MAP kinase. This kit was developed for and is recommended for immunohistochemistry only.

Source / Purification

Monoclonal antibody is produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr202/Tyr204 of human p44 MAP kinase.

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

• Kenerson, H. L. et al. (2002) Activated mammalian target of rapamycin pathway in the pathogenesis of tuberous sclerosis complex renal tumors. Cancer Res. 62, 5645-5650. This article references the use of Growth/Proliferation Marker: SignalStain® Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Detection Kit in the following applications: IHC-P (paraffin)
• Albanell, J. et al. (2001) Activated extracellular signal-regulated kinases: association with epidermal growth factor receptor/transforming growth factor alpha expression in head and neck squamous carcinoma and inhibition by anti-epidermal growth factor receptor treatments. Cancer Res. 61, 6500-6510. This article references the use of Growth/Proliferation Marker: SignalStain® Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Detection Kit in the following applications: IHC-P (paraffin)

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Companion Products

• 4376 Phospho-p44/42 MAPK (Thr202/Tyr204) (20G11) Rabbit mAb
• 4370 Phospho-p44/42 MAPK (Thr202/Tyr204) (D13.14.4E) Rabbit mAb
• 8103 SignalSlide™ Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Controls
• 1150 Phospho-p44/42 MAPK (Thr202/Tyr204) Blocking Peptide
• 9997 Tris Buffered Saline with Tween 20 (TBST - 10X)

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.