|Immunofluorescence (Immunocytochemistry)||1:50 - 1:200|
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.
NOTE: Formaldehyde is toxic, use only in a fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised November 2013
Protocol Id: 182
Monoclonal antibody is produced by immunizing animals with a protein fragment corresponding to residues near the carboxy terminus of human Fibronectin/FN1 protein.
Fibronectin is a multi-domain extracellular matrix (ECM) protein. The protein uses its different domains to bind distinct ECM components such as collagens, growth factors, and cell surface integrins to carry on its functions (1). Fibronectin has been implicated in many essential biological processes, including tissue repair, fibrosis, and tumor development (1,2). There are many fibronectin isoforms. Plasma fibronectin is synthesized by hepatocytes and exists as a compact, inactive conformation in the bloodstream. It is a major component of fibrin clots. Upon binding to integrins or other cell surface receptors, fibronectin switches to an extended conformation, and exposes its function domains to activated extracellular matrix assembly (3). Fibroblasts, endothelial cells, and many types of cancer cells have also been shown to synthesize cellular fibronectin isoforms. Among them, the EDA or EDB isoforms are particularly highly expressed during fibrosis and cancer development, and are potential diagnostic and therapeutic targets (4-6).
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