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F2006

Sigma-Aldrich

Fibronectin human plasma

lyophilized powder, BioReagent, suitable for cell culture

Synonym(s):
Fibronectin
EC Number:
MDL number:
eCl@ss:
42020141
NACRES:
NA.75

Quality Level

biological source

human plasma

sterility

sterile

product line

BioReagent

assay

≥85% (SDS-PAGE)

form

lyophilized powder

mol wt

450 kDa

packaging

pkg of 5 x 5 mg
pkg of 1 mg
pkg of 2 mg
pkg of 5 mg

application(s)

cell culture | mammalian: suitable

surface coverage

1‑5 μg/cm2

solubility

H2O: 1 mg/mL at 37 °C (Store reconstituted solution in working aliquots at -20°C or lower.)

UniProt accession no.

shipped in

dry ice

storage temp.

−20°C

Gene Information

human ... FN1(2335)

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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.

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 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

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.

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.

Storage Class Code

13 - Non Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US),Eyeshields,Gloves

Certificate of Analysis

Certificate of Origin

  1. 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.

  2. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  3. 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.

  4. How long can I store the fibronectin solution?

    The product is good for at least a year at 2-8 °C if kept sterile.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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. 

  12. 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.

  13. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Francois Bordeleau et al.
Proceedings of the National Academy of Sciences of the United States of America, 112(27), 8314-8319 (2015-06-25)
Alternative splicing of proteins gives rise to different isoforms that play a crucial role in regulating several cellular processes. Notably, splicing profiles are altered in several cancer types, and these profiles are believed to be involved in driving the oncogenic...
Kerim B Kaylan et al.
Integrative biology : quantitative biosciences from nano to macro, 8(12), 1221-1231 (2016-11-02)
Carcinoma progression is influenced by interactions between epithelial tumor cells and components of their microenvironment. In particular, cell-extracellular matrix (ECM) interactions are known to drive tumor growth, metastatic potential, and sensitivity or resistance to therapy. Yet the intrinsic complexity of...
Chia-Wen Chang et al.
Lab on a chip, 14(19), 3762-3772 (2014-08-07)
This paper reports a polydimethylsiloxane-polycarbonate (PDMS-PC) hybrid microfluidic device capable of performing cell culture under combinations of chemical and oxygen gradients. The microfluidic device is constructed of two PDMS layers with microfluidic channel patterns separated by a thin PDMS membrane....
Timothy E Quan et al.
Methods in molecular medicine, 135, 423-434 (2007-10-24)
Fibrocytes circulate in the peripheral blood, produce collagen and other matrix proteins, and express cell surface markers indicative of a hematopoetic origin distinguishing them from fibroblasts. Circulating fibrocytes were first identified in 1994 in a model system of wound repair...
Tiffany M Tsang et al.
Infection and immunity, 78(8), 3358-3368 (2010-05-26)
Yersinia pestis, the causative agent of plague, evades host immune responses and rapidly causes disease. The Y. pestis adhesin Ail mediates host cell binding and is critical for Yop delivery. To identify the Ail receptor(s), Ail was purified following overexpression...

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