Cell Signaling Technology

Product Pathways - Wnt / Hedgehog / Notch

Phospho-β-Catenin (Ser675) Antibody #9567

Applications Reactivity MW (kDa) Source
W IP H M R Mk (M) (C) (X) (Z) 92 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  C=Chicken  X=Xenopus  Z=Zebra Fish
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-β-Catenin (Ser675) Antibody detects endogenous levels of β-catenin only when phosphorylated at Ser675.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phosphopeptide (KLH-coupled) corresponding to residues surrounding Ser675 of human β-catenin. Antibodies are purified by peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from SK-N-MC cells, treated with forskolin (FSK) for 30 minutes or λ phosphatase for 1 hour, using Phospho-β-Catenin (Ser675) Antibody (upper) or β-Catenin Antibody (Amino-terminal Antigen) #9581 (lower).

Western Blotting

Western Blotting

COS-7 cells were transfected with cDNA encoding Flag-tagged wild type (WT) β-catenin or Ser-to-Ala β-catenin mutants as indicated. Cells were stimulated with 10 mM forskolin (FSK) for 5 minutes and lysed. β-catenin or its mutants were immunoprecipitated with Flag antibodies and analyzed by Western blotting using Phospho-β-Catenin (Ser552) Antibody #9566, Phospho-β-Catenin (Ser675) Antibody or a Flag antibody as indicated (Figures provided by Drs. Sebastien Taurin and Nickolai Dulin, Department of Medicine/Pulmonary, University of Chicago).

Background

β-catenin is a key downstream effector in the Wnt signaling pathway (1). It is implicated in two major biological processes in vertebrates: early embryonic development (2) and tumorigenesis (3). CK1 phosphorylates β-catenin on Ser45. This phosphorylation event primes β-catenin for subsequent phosphorylation by GSK-3 (4-6). GSK-3β destabilizes β-catenin by phosphorylating it at Ser33, Ser37 and Thr41 (7). Mutations in these phosphorylation sites, which result in the stabilization of β-catenin protein levels, have been found in many tumor cell lines (8).

PKA was shown to phosphorylate β-catenin at Ser675. Phosphorylation at Ser675 induces β-catenin accumulation in the nucleus and increases its transcriptional activity (9,10).

  1. Cadigan, K.M. and Nusse, R. (1997) Genes Dev. 11, 3286-3305.
  2. Wodarz, A. and Nusse, R. (1998) Annu. Rev. Cell. Dev. Biol. 14, 59-88.
  3. Polakis, P. (1999) Curr. Opin. Genet. Dev. 9, 15-21.
  4. Amit, S. et al. (2002) Genes Dev. 16, 1066-1076.
  5. Lin, C. et al. (2002) Cell 108, 837-847.
  6. Yanagawa, S. et al. (2002) EMBO J. 21, 1733-1742.
  7. Yost, C. et al. (1996) Genes Dev. 10, 1443-1454.
  8. Morin, P.J. (1997) Science 275, 1787-1790.
  9. Taurin, S. et al. (2006) J. Biol. Chem. 281, 9971-9976.
  10. Hino, S. et al. (2005) Mol. Cell Biol. 25, 9063-9072.

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