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F2006 Sigma

Fibronectin from human plasma

lyophilized powder, BioReagent, suitable for cell culture

Synonym: CIG, Cold insoluble globulin

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Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Components

Fibronectin is an adhesive glycoproteins found in both the cell surface and in plasma.

Application

Fibronectin from human plasma can be used with epithelial cells, mesenchymal cells, neuronal cells, fibroblasts, neural crest cells, and endothelial cells. It is recommended for use as a cell culture substratum at 1-5 μg/cm2 or 0.5-50 μg/mL. The optimal concentration does depend on cell type as well as the application and research objectives.

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 assays
• Protein specificity
• Immunofluorescence (used in staining of cell layers for membrane type-1 matrix metalloproteinase (MT1-MMP) for coating of slides)
• Extracellular matrix binding assay
• Used in cell migration study

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.

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.

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.

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.

Other Notes

Insolubles in our lyophilized fibronectin products is normal. Adding warm water carefully to the vial will, over time, provide a clear solution under a pellicle, a floating spider web-like material. Vials are filled with sufficient additional material to compensate for the loss of fibronectin in the pellicle. Protein concentration is tested in the reconstituted solution to ensure that the vials contain a minimum of the stated protein in a soluble form. Shaking or vortexing fibronectin in a liquid form will further denature the protein leading to decreased solubility.

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Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3

Frequently Asked Questions

Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
How do I choose which fibronectin product to use to coat culture surfaces for cell growth?
The species source for the fibronectin used when coating culture surfaces is not important.  It is the presence of the RGD binding site that is present in all fibronectins that promotes binding.
How long can I store the fibronectin solution?
The product is good for at least a year at 2-8 °C if kept sterile.
How long can I store fibronectin coated plates, etc?
Fibronectin coated cultureware can be stored for 2-4 weeks at 2-8 °C in a closed sterile container, or in sterile sealable bags.
What is the procedure for coating culture surfaces with fibronectin?
To coat culture surfaces: 1. Dilute fibronectin in sterile balanced salt solution and coat the culture surface (1-5 μg/cm2) with a minimal volume.  2. Allow to air dry for at least 45 minutes at room temperature.  Excess fibronectin may be removed by aspiration, but this is not necessary.
How do I solubilized Fibronectin?
Dissolve at 1 mg/mL in water. Allow to dissolve for at least 30 minutes at 37 °C. A small amount of undissolved material may remain. This will not affect product performance.
How do I test the wells/flasks that have been coated with fibronectin?
We test the coated culture surface by culturing cells.  We compare the adherence of the cells to wells that have been coated with media containing BSA (negative) and media containing 10% FBS (positive).  The cell growth should be not less than 50% of the positive control and greater than 50% of the negative control.
How do I determine how much fibronectin is in solution?
As the product may contain undissolved material, the amount of fibronectin in solution can be determined by UV absorbance.  The extinction coefficient for a 1% solution (10 mg/ml) is 13.5 at 280 nm.
How do I find price and availability?
There are several ways to find pricing and availability for our products.  Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers. 
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS. To access the shipping information for this material, use the link on the product detail page for the product, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
What is the difference between Fibronection Products F2006 and F1056?
The difference between Product Nos. F2006 and F1056 is in the purity specifications and endotoxin testing.  Product  No. F2006 has a purity specification of ≥90% and is not tested for endotoxin contamination.  Product No. F1056 has a purity specification of  ≥95% and is tested for endotoxin contamination with a specification of ≤100 EU/mg.
  What kind of solution is the A5955 - Antibiotic Antimycotic Solution (100×), Stabilized product suspended in?
A5955 - Antibiotic Antimycotic Solution (100×), Stabilized is supsended in a sodium citrate buffer solution. Source: Israel production group
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