Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-ASK1 (Ser83) Antibody #3761

Applications Reactivity Sensitivity MW (kDa) Source
W IP H Transfected Only 155 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human
Species cross-reactivity is determined by Western blot.

Protocols

Specificity / Sensitivity

Phospho-ASK1 (Ser83) Antibody detects transfected ASK1 only when phosphorylated at Ser83.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues around Ser83 of human ASK1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untransfected (lanes 1 and 4) or transfected wild type (lanes 2 and 5) or Ser83Ala mutant (lanes 3 and 6) ASK1, using Phospho-ASK1 (Ser83) Antibody (A and B) or ASK1 Antibody #3762 (C and D). The phospho-specificity of the antibody was further characterized by treating the membranes (B and D) with calf intestinal alkaline phosphatase (CIP) after Western transfer.

Background

Apoptosis signal-regulating kinase 1 (ASK1), a MAP kinase kinase kinase, plays essential roles in stress-induced apoptosis (1,2). ASK1 is activated in response to a variety of stress-related stimuli through distinct mechanisms and activates MKK4 and MKK3, which in turn activate JNK and p38 (3). Overexpression of ASK1 activates JNK and p38 and induces apoptosis in several cell types through signals involving the mitochondrial cell death pathway. Embryonic fibroblasts or primary neurons derived from ASK1-/- mice are resistant to stress-induced JNK and p38 activation and cell death (4,5). Phosphorylation at Ser967 is essential for ASK1 association with 14-3-3 protein and suppression of cell death (6). Oxidative stress induces dephosphorylation of Ser967 and phosphorylation of Thr845 in the activation loop of ASK1, and both are correlated with ASK1 activity and ASK1-dependent apoptosis (7,8). On the other hand, Akt phosphorylates ASK1 at Ser83, which attenuates ASK1 activity and promotes cell survival (9).

ASK1 is a substrate for phosphorylation at Ser83 by Akt, and that this phosphorylation is associated with a decrease in stimulated ASK1 kinase activity (6).

  1. Ichijo, H. et al. (1997) Science 275, 90-94.
  2. Wang, X.S. et al. (1996) J. Biol. Chem. 271, 31607-31611.
  3. Matsuzawa, A. and Ichijo, H. (2001) J. Biochem. (Tokyo) 130, 1-8.
  4. Tobiume, K. et al. (2001) EMBO Rep. 2, 222-228.
  5. Nishitoh, H. et al. (2002) Genes Dev. 16, 1345-1355.
  6. Zhang, L. et al. (1999) Proc. Natl. Acad. Sci. USA 96, 8511-8515.
  7. Tobiume, K. et al. (2002) J. Cell. Physiol. 191, 95-104.
  8. Goldman, E.H. et al. (2004) J. Biol. Chem. in press, .
  9. Kim, A.H. et al. (2001) Mol. Cell. Biol. 21, 893-901.

Application References

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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