Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-Erk5 (Thr218/Tyr220) Antibody #3371

Applications Reactivity Sensitivity MW (kDa) Source
W H M R Transfected Only 115 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-Erk5 (Thr218/Tyr220) Antibody detects immunoprecipitated or transfected levels of Erk5 phosphorylated at threonine 218 and tyrosine 220. This antibody cross-reacts with phosphorylated Erk1 and Erk2. It does not cross-react with phosphorylated p38 MAPK or SAPK/JNK.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr218/Tyr220 of human Erk5. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Immunoprecipitation of Erk5 from NIH/3T3 cells, untreated or PDGF-treated, using Erk5 Antibody #3372, followed by Western blot with Phospho-Erk5 (Thr218/Tyr220) Antibody #3371, or Erk5 Antibody #3372.

Background

ERK5 (Mitogen-activated protein kinase 7, Big mitogen-activated protein kinase 1) is a member of the MAPK superfamily implicated in the regulation numerous cellular processes including proliferation, differentiation, and survival (1,5-7). Like other MAPK family members, ERK5 contains a canonical activation loop TEY motif (Thr218/Tyr220) which is specifically phosphorylated by MAP2K5 (MEK5) in a growth factor-dependent, Ras-independent mechanism (2-4). For example, EGF stimulation promotes ERK5 phosphorylation which induces its traslocation to the nucleus where it phosphorylates MEF2C and other transcriptional targets (2,3). ERK5 is also activated in response to granulocyte colony-stimulating factor (G-CSF) in hematopoetic progenitor cells where it promotes survival and proliferation (4). In neuronal cells, ERK5 is required for NGF-induced neurite outgrowth, neuronal homeostatis, and survival (11,12). ERK5 is thought to play a role in blood vessel integrity via maintainence of endothelial cell migration and barrier function (8-10). Although broadly expressed, research studies have shown that mice lacking erk5 display numerous cardiac defects, suggesting ERK5 plays a critical role in vascular development and homeostasis (1,5).

  1. Zhou, G. et al. (1995) J Biol Chem 270, 12665-9.
  2. Kato, Y. et al. (1998) Nature 395, 713-6.
  3. Kato, Y. et al. (1997) EMBO J 16, 7054-66.
  4. Dong, F. et al. (2001) J Biol Chem 276, 10811-6.
  5. Hayashi, M. and Lee, J.D. (2004) J Mol Med 82, 800-8.
  6. Wang, X. and Tournier, C. (2006) Cell Signal 18, 753-60.
  7. Nishimoto, S. and Nishida, E. (2006) EMBO Rep 7, 782-6.
  8. Spiering, D. et al. (2009) J Biol Chem , .
  9. Sawhney, R.S. et al. (2009) J Cell Physiol 219, 152-61.
  10. Zhao, Z. et al. (2009) Mol Cell Biochem 322, 171-8.
  11. Obara, Y. et al. (2009) J Biol Chem , .
  12. Finegan, K.G. et al. (2009) Cell Death Differ 16, 674-83.

Application References

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Companion Products

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

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