Semi-purified Vinculin. Cellular Localization: Cell membrane and cytoplasmic. MW of Antigen: 130kD (vinculin) and 150kD (meta-vinculin).
Vinculin is an attachment protein involved in the indirect binding of intracellular actin filaments to extracellular fibronectin. Vinculin binds to actin capping protein and alpha actinin, which in turn binds to actin. The binding of fibronectin is also indirect where vinculin binds to talin, which in turn binds to the intracellular domain of the fibronectin receptor which itself is the ligand for fibronectin. Vinculin is widely distributed in tissue and is expressed where smooth muscle actin and fibroblasts attach to the extracellular matrix. Applications: Suitable for use in Immunofluorescence, Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications not tested. Recommended Dilutions: Western Blot: 1-2ug/ml for 2 hours at RT. Immunoprecipitation (Denatured): 2ug/mg protein lysate (Use Protein G). Immunohistochemistry (Formalin/paraffin): 1:50 for 30 minutes at RT. Staining of formalin/paraffin tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0, for 10-20 minutes followed by cooling at RT for 20 minutes. Optimal dilutions to be determined by the researcher. Positive Control: HeLa cells. Skin Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
