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Technical Data Sheet


β-Catenin Tyr-142 & Tyr-654 Phosphorylation
Immunocytochemistry Kit


Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Product References
CP1081 Tsuneki, M. et al. (2014) Mol Cell Biol. 34(24): 4485 IF: mouse EOMA, brain endothelial cells
CP4021 Lu, Y. et al.(2014) Stem Cells Dev. 23(15):1755. IF: chicken embryonic fibroblasts
CP1081 Qi, F. et al. (2013) Am J Pathol. 183(5):1654. WB: human mesothelial
CP4021 Condello, S. et al. (2013) FASEB Journal 27(8): 3100. WB: ovarian cancer cells
CM1181 Condello, S. et al. (2013) FASEB J 27(8):3100. WB & IHC: Ovarian Cancer Cells
CM1181 Yakubov, B. et al. (2013) Neoplasia 15(6):609. WB: Human Ovarian Cancer Cells
CP1081 Krejci, P. et al. (2012) PLoS ONE. 7(4):e35826. WB: rat chondrosarcoma cells
CP1081 Beard, R.S. et al. (2011) Blood. 118(7):2007. ELISA: mouse microvascular ECs
Kit Summary
The β-Catenin phospho-regulation kit can be used for immunocytochemical co-localization of β-Catenin phosphorylation at Tyr-142 and Tyr-654 relative to total β-Catenin. The kit includes rabbit polyclonal and mouse monoclonal antibodies, along with goat-anti-rabbit conjugated to DyLight® 594 and goat anti-mouse conjugated to DyLight® 488 for dual labeling experiments.

Immunocytochemical labeling of phosphorylated β-Catenin in paraformaldehyde-fixed and NP-40-permeabilized rabbit spleen fibroblasts (top row) and A431 cells (bottom row). The fixed cells were labeled with mouse monoclonal antibody to β-Catenin (CM1811) (top and bottom left), and phospho-specific β-Catenin (Tyr-142) (top right) or β-Catenin (Tyr-654) (bottom right). The antibodies were detected using Goat anti-Mouse DyLight® 488 or Goat anti-Rabbit DyLight® 594.

β-Catenin is a 92 kDa protein that binds to the cytoplasmic tail of E-Cadherin. The cadherins, transmembrane adhesion molecules, are found with catenins at adherens junctions. Deletions in the cytoplasmic domain of E-Cadherin eliminate catenin binding and result in a loss of cell adhesion. Tyrosine phosphorylation of β-Catenin can regulate its interaction with critical components of adherens junctions. Both Fer and Fyn Kinases phosphorylate tyrosine 142 in vitro. Overexpression of these kinases in epithelial cells disrupts interactions between α- and β-Catenins. The phosphorylation of tyrosine 142 may act as a switch from the transcriptional to the adhesive role of β-Catenin. Src family kinases can also phosphorylate tyrosine 86 and 654 in β-Catenin. The Tyr-654 phosphorylation regulates β-Catenin binding to E-cadherin. Thus, site-specific tyrosine phosphorylation of β-Catenin may regulate protein-protein interactions, leading to changes in cell adhesion.

Background References
Ozawa, M. et al. (1990) Proc. Natl. Acad. Sci. USA 87:4246.
Roura, S. et al. (1999) J Biol Chem. 274(51) :36734.
Piedra, J. et al. (2003) Mol. Cell. Biol. 23(7):2287.
Brembeck, F.H. et al. (2004) Genes Dev. 18(18):2225.
Rhee, J. et al. (2007) Nat. Cell Biol. 9(8):883.
Buffer and Storage
Mouse monoclonal, rabbit polyclonal, and secondary reagents are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
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