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#7278Store at 4C and -20C

 

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Species Cross Reactivity

H M

UniProt ID:

#P27361

Entrez-Gene Id:

#5595

Cell Signaling Technology

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For Research Use Only. Not for Use in Diagnostic Procedures.
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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.

Reagents not supplied

Phosphate Buffered Saline (PBS-20X) #9808 Phosphate Buffered Saline with Tween -20 (PBST-20X) #9809 Cell Lysis Buffer (10X) #9803 TMB Substrate #7004 STOP Solution #7002 Blocking Buffer- PBS+0.05% Tween-20, 1% BSA 96 Well Microplates** Microplate Reader ** Antibody Pairs have been validated on Corning® 96 Well Clear Polystyrene High Bind Stripwell™ Microplates (#2592) and Corning® 96 Well EIA/RIA Easy Wash™ Clear Flat Bottom Polystyrene High Bind Microplates (#3369).

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.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

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    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PathScan is a registered trademark of Cell Signaling Technology, Inc.
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