Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-ASK1 (Ser967) Antibody #3764

Applications Reactivity Sensitivity MW (kDa) Source
W H (M) (R) Transfected Only 155 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-ASK1 (Ser967) Antibody detects transfected ASK1 only when phosphorylated at serine 967.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around serine 967 of human ASK1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS7 cells transfected with wild-type or Ser967Ala mutant ASK1, using Phospho-ASK1 (Ser967) Antibody (A,B) or ASK1 Antibody #3762 (C,D). The phospho-specificity of the antibody was further characterized by treating the membrane with calf intestinal alkaline phosphatase (CIP) after Western transfer (B,D).

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 as well as cell death (4,5). Phosphorylation at Ser967 is essential for ASK1 association with 14-3-3 proteins and suppression of cell death (6). Oxidative stress induces dephosphorylation of Ser967 and phosphorylation of Thr845 in the activation loop of ASK1, both of which are correlated with ASK1 activity and ASK1-dependent apoptosis (7,8). Akt phosphorylates ASK1 at Ser83, which attenuates ASK1 activity and promotes cell survival (9).

Phosphorylation at Ser967 of ASK1 leads to association of 14-3-3 and suppression of 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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For Research Use Only. Not For Use In Diagnostic Procedures.

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