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Technical Data Sheet

CK6310

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N-Cadherin Phospho-Regulation
Antibody Sampler Kit
Price

$395

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
CP1951 Wang, M. et al. (2014) Cell Mol Neurobiol. 34(1):123 WB: MN9D neurons
CP1751 Chang, Y.J. et al. (2014) Mol Cell Biol. 34(6):1003 WB: PC12-SH2B1 cell
CP1751 Wu, Y. et al. (2012) Int J Oncol. 41(4):1337. WB: human pancreatic cancer cells (PANC-1 PaCa-2)
CP1751 Sato, F. et al. (2012) Intern J Molec Med 30:495. WB: Human pancreatic cancer BxPC-3 cells
CM1701 Wang, T.C. et al. (2011) J. Cell. Physiol. 226:2063. WB, ICC: rat PC12 cells
CP1751 Ferreri, D.M. et al. (2008) Cell Comm Adh. 15(4):333. WB: human, bovine VECs
Kit Summary
The N-cadherin phospho-regulation antibody sampler kit can be used to examine phosphorylation of N-cadherin at Tyr-820 and Tyr-860. The kit also includes monoclonal and polyclonal antibodies to monitor total expression levels of N-Cadherin.
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Western blot image of human endothelial cells untreated (lanes 1 & 3) or treated with pervanadate (1 mM) for 30 min (lanes 2, 4, 5 & 6). The blots were probed with anti-N-Cadherin (Cytoplasmic) (lanes 1 & 2) and anti-N-cadherin (Tyr-820) (lanes 3-6). The latter antibody was used in the presence of no peptide (lane 4), phospho-N-cadherin (Tyr-820) peptide (lane 5), or phospho-N-cadherin (Tyr-860) peptide (lane 6).

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Immunocytochemical labeling of phosphorylated N-Cadherin in pervanadate-treated mouse C2C12. The cells were probed with N-Cadherin (Cytoplasmic) and N-Cadherin(Tyr-860) antibodies, followed by immunofluorescent detection.

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Background
Cadherins are transmembrane glycoproteins vital in calcium-dependent cell-cell adhesion during tissue differentiation. Cadherins cluster to form foci of homophilic binding units. A key determinant to the strength of the cadherin-mediated adhesion may be by the juxtamembrane region in cadherins. This region induces clustering and also binds to the protein p120 catenin. The cytoplasmic region is highly conserved in sequence and has been shown experimentally to regulate the cell-cell binding function of the extracellular domain of E-cadherin, possibly through interaction with the cytoskeleton. Many cadherins are regulated by phosphorylation, including N-cadherin and E-cadherin. N-cadherin is phosphorylated by c-Src at Tyr-820, Tyr-853, Tyr-860, Tyr-884, and Tyr-886. Phosphorylation of Tyr-860 can disrupt cadherin binding to β-catenin. Since many of these tyrosine sites are conserved in the cadherin family, phosphorylation of these sites may be critical for cadherin function.

Background References
Xu, Y. et al. (1997) J. Biol. Chem. 272(21):13463.
Xu, Y. & Carpenter, G. (1999) J. Cell. Bioch. 75:264.
Qi, J. et al. (2006) Mol. Biol. Cell 17(3):1261.
Buffer and Storage
Rabbit polyclonal and mouse monoclonal antibodies 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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