Product Pathways - MAPK Signaling
p44/42 MAP Kinase Assay Kit (Nonradioactive) #9800
| Kit Includes | Quantity | Source |
|---|---|---|
| Immobilized Phospho-p44/42 MAPK (Thr202/Tyr204) Mouse mAb # 9109 | 600 microliters | Mouse |
| Phospho-Elk-1 (Ser383) Antibody # 9181 | 100 microliters | Rabbit |
| Elk-1 Fusion Protein # 9184 | 80 micrograms | |
| Kinase Buffer (10X) # 9802 | 15 milliliters | |
| Cell Lysis Buffer (10X) # 9803 | 15 milliliters | |
| ATP (10 mM) # 9804 | 50 microliters | |
| Anti-rabbit IgG, HRP-linked Antibody # 7074 | 50 microliters | Goat |
| Anti-biotin, HRP-linked Antibody # 7075 | 100 microliters | Goat |
| Biotinylated Protein Ladder Detection Pack # 7727 | 100 microliters | |
| 20X LumiGLO® Reagent and 20X Peroxide # 7003 | 5 milliliters each | |
| Active p42 MAP Kinase (10 ng/?l) | 10 microliters |
Molecular Weight
45
Reactivity
H M R
Reactivity Key: H=Human M=Mouse R=Rat
Description
Nonradioactive p44/42 MAP Kinase Assay Kit provides all the reagents necessary for measuring p44/42 MAP kinase activity in the cell. Immobilized Phospho-p44/42 MAPK (Thr202/Tyr204) mAb is used to immunoprecipitate active p44/42 MAP kinase from cell extracts, then an in vitro kinase assay is performed using Elk-1 protein as a substrate. Elk-1 phosphorylation is then detected by Western blotting using Phospho-Elk-1 (Ser383) Antibody.Improvements Over Conventional Assays:Improved sensitivity without radioactivityPhospho-specific antibodies allow site-specific analysisDramatically increased signal to noise ratio over conventional IP/kinase assaysLow background activityIncludes complete system to assay kinase activity
Improvements Over Conventional Assays
- No Radioactivity
- Improved sensitivity without radioactivity.
- Specificity
- Phospho-specific antibodies allow site-specific analysis.
- Signal to Noise
- Dramatically increased signal to noise ratio over conventional IP/kinase assays.
- Low Background Activity
- Complete System
- Includes everything needed to assay kinase activity.
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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- Baccarini, M. (2005) FEBS Lett 579, 3271-7.
- Meloche, S. and Pouyssegur, J. (2007) Oncogene 26, 3227-39.
- Roberts, P.J. and Der, C.J. (2007) Oncogene 26, 3291-310.
- Rubinfeld, H. and Seger, R. (2005) Mol Biotechnol 31, 151-74.
- Murphy, L.O. and Blenis, J. (2006) Trends Biochem Sci 31, 268-75.
- Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
- Marais, R. et al. (1993) Cell 73, 381-93.
- Kortenjann, M. et al. (1994) Mol Cell Biol 14, 4815-24.
- Owens, D.M. and Keyse, S.M. (2007) Oncogene 26, 3203-13.
Application References
- Russell, K.S. et al. (2000) Proc Natl Acad Sci U S A 97, 5930-5. This article references the use of p44/42 MAP Kinase Assay Kit (Nonradioactive) in the following applications: In vitro kinase assay
- Tan, J. et al. (2000) J Neurosci 20, 7587-94. This article references the use of p44/42 MAP Kinase Assay Kit (Nonradioactive) in the following applications: In vitro kinase assay
- Jost, M. et al. (2001) J Biol Chem 276, 6320-6. This article references the use of p44/42 MAP Kinase Assay Kit (Nonradioactive) in the following applications: In vitro kinase assay
- Skapenko, A. et al. (2001) J Immunol 166, 4283-92. This article references the use of p44/42 MAP Kinase Assay Kit (Nonradioactive) in the following applications: In vitro kinase assay
- Gui, Y. and Zheng, X.L. (2003) J Biol Chem 278, 53017-25. This article references the use of p44/42 MAP Kinase Assay Kit (Nonradioactive) in the following applications: In vitro kinase assay
- Hsu, H.Y. et al. (2004) J Immunol 173, 5989-99. This article references the use of p44/42 MAP Kinase Assay Kit (Nonradioactive) in the following applications: In vitro kinase assay
- Wong, C.H. et al. (2005) Endocrinology 146, 1192-204. This article references the use of p44/42 MAP Kinase Assay Kit (Nonradioactive) in the following applications: In vitro kinase assay
- He, Z. et al. (2005) J Biol Chem 280, 2446-54. This article references the use of p44/42 MAP Kinase Assay Kit (Nonradioactive) in the following applications: In vitro kinase assay
- Schlezinger, J.J. et al. (2006) Toxicol Sci 92, 433-44. This article references the use of p44/42 MAP Kinase Assay Kit (Nonradioactive) in the following applications: In vitro kinase assay
- Jia, W. et al. (2006) Mol Cancer Res 4, 549-62. This article references the use of p44/42 MAP Kinase Assay Kit (Nonradioactive) in the following applications: In vitro kinase assay
- Zampieri, C.A. et al. (2007) J Virol 81, 1230-40. This article references the use of p44/42 MAP Kinase Assay Kit (Nonradioactive) in the following applications: In vitro kinase assay
- Ha, U. et al. (2007) J Immunol 178, 1736-47. This article references the use of p44/42 MAP Kinase Assay Kit (Nonradioactive) in the following applications: In vitro kinase assay
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Companion Products
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- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack