Anti-phospho-Vinculin (Tyr1065)

Vinculin is an ubiquitously expressed cytoskeletal protein (~130kD) involved in cell adhesion and cell migration. The vinculin protein consists of a globular head domain connected to an elongated tail region by a proline-rich domain. The head region contains binding sites for two cytoskeletal proteins, a-actinin and talin, as well as a binding site for the tail region of vinculin itself. The tail region contains binding sites for actin, the cytoskeletal protein, paxillin, and PI(4,5)P2. In the inactive state the head region of vinculin is bound to the tail region, resulting in inaccessibility of the other protein binding sites. Binding of PI(4,5)P2 releases the head-tail interaction allowing binding of other proteins to vinculin. These regulatory events play an important role in the formation, maintenance, and breakdown of focal adhesions that occur during cell adhesion and migration. In addition to these protein binding sites, the head and tail regions of vinculin have multiple potential phosphorylation sites. Phosphorylation of vinculin at pY1065 is c-Src dependent. This has been observed in vitro and in activated platelets. Phosphorylation of vinculin on tyrosine residue 1065 has been shown to affect head-tail interactions and play a role in cell spreading, however, it does not affect the interaction between vinculin and actin or cellular localization. Applications: Suitable for use in Western Blot following Immunoprecipitation. Other applications not tested. Recommended Dilution: Western Blot: 1:1000 , Optimal dilutions to be determined by the researcher. Positive Control: COS cells co-transfected with activated Src and His-tagged chicken vinculin cDNA were treated with vanadate for 24 hr. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after reciept. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.