Description
General description
Fibronectins (FNs) are high-molecular-mass adhesive glycoproteins of the extracellular matrix (ECM) and body fluids. It is a dimer composed of two subunits linked by two disulfide bonds. Each monomer of FN consists of three types of repeating units: 12 FN repeats of type I (approximately 40 amino acids each), two type II repeats (around 60 amino acids), and 15-17 type III repeats (approximately 90 amino acids). These repeating units contribute to the structural and functional diversity of fibronectin molecules.
Application
Human fibronectin (HFN) is suitable for use as an attachment factor in the propagation of cells in vitro when used to coat cell culture surfaces, including plastic ware, glassware, and microcarrier beads.
Biochem/physiol Actions
Fibronectin plays a crucial role in mediating diverse cellular interactions and is involved in various processes, including cell adhesion, the establishment and maintenance of cell morphology, cell migration, growth, and differentiation. It interacts with several extracellular matrix (ECM) and cell surface proteins, such as collagen, heparin, fibrin, and specific cell membrane receptors. Fibronectin can also serve as a ligand for multiple integrins, including the well-known fibronectin receptor alpha5-beta1, which binds to the arginylglycylaspartic acid (RGD) sequence in the repeat III-10 region of fibronectin. This extensive network of interactions highlights fibronectin′s significance in regulating cellular behavior and mediating tissue function.
Physical form
Liquid in 50 mM tris-buffered saline (TBS), pH 7.5,containing no preservatives.Lyophilized from 50 mM tris-buffered saline (TBS), pH 7.5,containing no preservatives (100 MG pack size). Reconstitute the entire vial ofhFN to 1 mg/mL using a sterile balanced salt solution at 37°C.
Preparation Note
Maintain at 2-8°C for up to 6 months from date of receipt. Do not freeze.
Analysis Note
A double band of 220 kDa is present under reduced conditions.hFN is purified by affinity chromatography on gelatin agarose, followed by chromatography on heparin-agarose.Plasma from donors has been screened, and shown to be negative for HIV, HTLV, Hepatitis B and C.
Other Notes
Suggested Procedure for Coating Cell Cultureware
1.Determine the amount of HFN needed to coat culture vessels by multiplying the total surface area (cm2) by the desired concentration (μg/mL) of HFN. Recommended amount is 2-10 μg/cm2.
2.Wet the surface of each culture vessel to be coated with a minimum amount of sterile balanced salt solution (serum and protein free) required to cover the entire area.
3.Introduce the proper CO2 atmosphere, if required.
4.Add the calculated amount of HFN to each culture vessel.
5.Allow HFN to adsorb to the surface of the vessel for 5-20 minutes.
6.Remove residual balanced salt solution before proceeding with standard cell culture procedures
Product is filtered using a 0.1 micron filter during production. It is recommended to filter product again before use/after reconstitution.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
RESEARCH USE ONLY. This product is regulated in France when intended to be used for scientific purposes, including for import and export activities (Article L 1211-1 paragraph 2 of the Public Health Code). The purchaser (i.e. enduser) is required to obtain an import authorization from the France Ministry of Research referred in the Article L1245-5-1 II. of Public Health Code. By ordering this product, you are confirming that you have obtained the proper import authorization.
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
