Cell Signaling Technology

Product Pathways - MAPK Signaling

p44/42 MAPK (Erk1/2) Antibody #9102

Applications Reactivity Sensitivity MW (kDa) Source
W IP IHC-P IF-IC F H M R Hm Mk Mi Z B Pg Sc Endogenous 42, 44 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  Mi=Mink  Z=Zebrafish  B=Bovine  Pg=Pig  Sc=S. cerevisiae
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

p44/42 MAPK (Erk1/2) Antibody detects endogenous levels of total p44/42 MAP kinase (Erk1/Erk2) protein. In some cell types, this antibody recognizes p44 MAPK more readily than p42 MAPK. The antibody does not recognize either JNK/SAPK or p38 MAP kinase.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to a sequence in the C-terminus of rat p44 MAP Kinase. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells transfected with 100 nM control siRNA #6201 (-) or p44 MAPK (Erk1) siRNA (+), using p44/42 MAPK (Erk1/2) Antibody #9102 and GCK Antibody #3782. The Erk1/2 antibody confirms silencing of Erk1 expression, and GCK Antibody is used to control for loading and specificity of p44 MAPK (Erk1) siRNA.

Western Blotting

Western Blotting

Western blot analysis of extracts from serum-induced PC12 cells, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody #9101 (upper) or control p44/42 MAPK (Erk1/2) Antibody (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear and cytoplasmic localization, using p44/42 MAPK (Erk1/2) Antibody.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, using p44/42 MAPK (Erk1/2) Antibody (blue) compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent images of C6 cells serum-starved (left) and serum-treated (center), labeled with p44/42 MAPK (Erk1/2) Antibody (green) compared to an isotype control (right). Actin filaments have been labeled with Alex Fluor® 555 phalloidin. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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

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

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