Cell Signaling Technology

Product Pathways - Wnt / Hedgehog / Notch

β-Catenin Antibody #9562

Applications Reactivity Sensitivity MW (kDa) Source
W IP IHC-P ChIP H M R Mk (Z) Endogenous 92 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Z=Zebrafish
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

β-Catenin Antibody detects endogenous levels total of β-catenin. This antibody does cross-react with endogenous levels of γ-catenin.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues around Ser37 of human β-catenin. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293, HeLa, NIH/3T3 and C6 cells using β-Catenin Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from SW480 cells using β-Catenin Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using beta-Catenin Antibody.


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HCT116 cells and either 20 μl of β-Catenin Antibody or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using using SimpleChIP® Human CaMK2D Intron 3 Primers #5111, human c-Myc promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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 at 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 at these sites result in the stabilization of β-catenin protein levels and have been found in many tumor cell lines (8).

  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.

Application References

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Companion Products

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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