Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-p44 MAPK (Thr202/Tyr204) Sandwich ELISA Antibody Pair #7278

Kit Includes Volume Cap Color
p44 MAPK Capture Ab (100X) 0.4 ml Pink
P-p44/42 MAPK (T202/ Y204) Detection Ab (100X) 0.4 ml Blue
Anti-mouse IgG, HRP-linked Antibody 0.04 ml Yellow

Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.

Species Cross-Reactivity

H M

Reactivity Key:  H=Human  M=Mouse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Description

CST's PathScan® Phospho-p44 MAPK (Thr202/Tyr204) Sandwich ELISA Antibody Pair is offered as an alternative to our PathScan® Phospho-p44 MAPK (Thr202/Tyr204) Sandwich ELISA Kit, #7315. Capture and detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are provided for performing 4 x 96 well ELISAs. p44 MAPK Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added, followed by a Phospho-p44/42 MAPK (Thr202/Tyr204) Detection Antibody and HRP-conjugated secondary antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance at 450 nm is proportional to the quantity of phospho-p44/42 MAPK (Thr202/Tyr204) protein.Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Sandwich ELISA

Sandwich ELISA

The relationship between lysate protein concentration from untreated and PDGF-treated NIH/3T3 cells and the absorbance at 450 nm using PathScan® Phospho-p44 MAPK (Thr202/Tyr204) Sandwich ELISA Antibody Pair #7278 is shown. NIH/3T3 cells were treated with PDGF for 5 minutes at 37ºC 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 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.

  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

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

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

Products