Application

Fibronectin from human plasma is used for the following applications:Used in cell seeding and neuronal differentiation (as one of the components in expansion medium for the re-suspension of CBSCs (Cord blood stem cells) and is also used in differentiation medium)Used in cell culture (the culture chamber was coated with fibronectin for the expression of HEK293 cells)Used in murine embroyonic stem cell line (CCE) culture and induction of endothelial cell differentiation (for the induction of endothelial cell differentiation, cultured cells were pre-coated with fibronectin)In-vitro fibronectin (FN) pulldown assaysProtein specificityImmunofluorescence (used in staining of cell layers for membrane type-1 matrix metalloproteinase (MT1-MMP) for coating of slides)Extracellular matrix binding assayUsed in cell migration study

Biochem/physiol Actions

Fibronectin (FN) plays an essential role in multiple biological functions such as, angiogenesis, cell migration and differentiation and wound healing. Studies in Chinese Han population show that mutations in this gene might be linked with susceptibility to knee osteoarthritis (OA). Splice variant extra domain-B (EDB) of FN is a hallmark of tumor angiogenesis, and with increased matrix stiffness there is an increase in the production of this isoform by endothelial cells. In colorectal cancer (CRC), EDA is linked with metastasis, vasculogenesis and tumorigenesis and promotes epithelial-mesenchymal transition.

Caution

The reconstituted solution should be stored in working aliquots at -20°C or lower. Vortexing, excessive agitation, or additoinal freezing and thawing of reconstituted fibronectin are not recommended.

General description

Fibronectin (FN) is a multifunctional glycoprotein, the gene of which is localized to human chromosome 2q34-36. This gene spans 75kb and contains 50 exons. It exists as two major isoforms, one soluble and one insoluble form. The former is present in plasma, whereas the insoluble form resides in tissues and extracellular matrix (ECM) of cartilage. There are around 20 different forms of FN due to alternative splicing of a variable region and two type III exons called Extra Domains A and B.

Preparation Note

This product is lyophilized from 0.05 M Tris buffered saline at pH 7.5. It is soluble at 1 mg/mL in water, and should be allowed to dissolve for at least 30 minutes at 37°C. A small amount of undissolved material may remain, but it will not affect performance. The protein homogeneity is evaluated by immunoelectrophoresis, is endotoxin tested, and the source material was tested negative for HIV, hepatitis B and hepatitis C. In coating culture surfaces, fibronectin should be diluted in sterile balanced sterile salt solution and coated with minimal volume. Surface should then be air dried for 45 minutes at room temp and can be stored for 2-4 weeks at 2-8°C.