Smoothelin Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Smoothelin is a constituent of the smooth muscle cell (SMC) cytoskeleton. Antibodies directed to smoothelin are a useful tool to monitor SMC differentiation. Smoothelin is exclusively expressed in fully differentiated (contractile) SMCs. RNA and protein analyses reveal a broad species distribution of this protein. Cells with SMC like characteristics, such as myofibroblasts, myoepithelial cells, and skeletal and cardiac muscle do not contain smoothelin. Confocal scanning laser microscopy of tissue sections, as well as transfection studies, show a filamentous organization of smoothelin that is different from that of desmin and vimentin. Smoothelin colocalizes with actin stress fibers. Two tissue specific isoforms have been identified: a 59 kDa isoform specific for visceral SMC; and a 100 kDa specific for vascular SMC. Human smoothelin is encoded by a single copy gene which is located on chromosome 22. The distribution of smoothelin positive cells indicates a correlation between smoothelin expression and smooth muscle tissue contractility. Smoothelin is a useful tool in the evaluation of atherosclerotic lesions.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-05688

• Smoothelin Antibodies
• Smoothelin ELISA Kits
• Smoothelin Lysates
• Smoothelin Proteins