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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 30 min at room temperature. A suggested positive control is breast carcinoma.
Paxillin is a focal adhesion protein and a substrate for several tyrosine kinases such as src, FAK, and p210BRC/ABL. The tyrosine phosphorylation of paxillin is affected by conditions that change cell-cell adhesion. Paxillin associates tightly with FAK and Crk through its SH2 domain. This interaction is independent of the extracellular matrix. Although paxillin was initially discovered in fibroblasts, its phosphorylation may also be important during neurite extension during differentiation.
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