Cell Signaling Technology

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PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Antibody Pair #7246

Kit Includes Volume Cap Color
Phospho-p44/42 MAPK (Thr202/Tyr204) Rabbit Capture Antibody (100X) 0.4 milliliters Pink
p44/42 MAPK Mouse Detection Antibody (100X) 0.4 milliliters Blue
Anti-Mouse IgG, HRP-Linked Antibody (1000X) 0.04 milliliters Yellow

Capture and Detection Antibodies are stored at 4°C. HRP-Linked Secondary Antibody is stored at -20°C.

Species Cross-Reactivity

H M

Reactivity Key:  H=Human  M=Mouse

Description

CST's PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Kit #7177. Capture and detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Phospho-p44/42 MAPK Rabbit Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysate is added followed by a p44/42 MAPK Mouse Detection Antibody and an HRP-conjugated Anti-Mouse IgG Antibody. HRP substrate (TMB) is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of p44/42 MAPK phosphorylated at Thr202/Tyr204.*Antibodies in this kit are custom formulations specific to the kit.

Sandwich ELISA

Sandwich ELISA

The relationship between the protein concentration of the lysate from untreated and PDGF-treated NIH/3T3 cells and the absorbance at 450 nm using PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Antibody Pair #7246 is shown. NIH/3T3 cells were treated with PDGF (100 ng/mL) for 5 minutes at 37°C and then lysed.

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.

  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.

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