Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-ASK1 (Thr845) Antibody #3765

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

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse
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 (Thr845) Antibody detects transfected ASK1 only when phosphorylated at threonine 845.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr845 of mouse ASK1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untransfected (lane 1), or transfected with wild-type ASK1 (lanes 2,3), using Phospho-ASK1 (Thr845) Antibody (upper) or ASK1 Antibody #3762 (lower). In lane 3, the lysate was treated with lamda phosphatase to demonstrate phospho-specificity of Phospho-ASK1 (Thr845) Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from HEK293 cells, untransfected (lane 1) or transfected with wild-type ASK1 (lane 2) or T845A mutant ASK1 (lane 3), using Phospho-ASK1 (Thr845) Antibody (upper) or ASK1 Antibody #3762 (lower). (Lysates were provided by Dr. Wang Min, Dept. of Pathology, Yale University, New Haven, CT).

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).

  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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