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Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 10 min at room temperature. A suggested positive control is skin, tonsil or squamous epithelium.
N-cadherin is a 140 kDa protein belonging to a family of transmembrane molecules that mediate calcium-dependent intercellular adhesion. Cadherins are involved in controlling morphogenetic movements during development and regulate cell surface adhesion through homotypic adhesion with the same cadherin species. N-cadherin's function is dependent on its association with the actin-cytoskeleton and is mediated through interactions between the C-terminal region of N-cadherin and the cytoplasmic catenin proteins. The stability of this association is regulated by phosphorylation and dephosphorylation of beta-catenin. Further, N-cadherin is a calcium dependent cell-cell adhesion glycoprotein comprised of five extracellular cadherin repeats, a transmembrane region and a highly conserved cytoplasmic tail. The protein functions during gastrulation and is required for establishment of left-right asymmetry.
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