Recombinant Human Fibronectin, ACFP Protein

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human Fibronectin, ACFP Protein Summary

• Details of Functionality: Measured by its ability to support cell attachment and spreading when used as a substratum for cell culture. In this application, the recommended concentration for this effect is typically 1-5 μg/cm².
  Fibronectin can also be added to the media to support cell spreading at a concentration of 0.5-50 μg/mL.
  Optimal concentrations will need to be determined for individual user applications.
• Source: Spodoptera frugiperda, Sf 9 (baculovirus)-derived human Fibronectin protein
  Met1-Pro1908, with a C-terminal 6-His tag
  Produced in an animal component free process (ACFP).
• Accession #: CAE45932
• N-terminal Sequence: No results obtained. Gln32 inferred from enzymatic pyroglutamate treatment revealing Ala33
• Protein/Peptide Type: Animal Component Free Recombinant Proteins
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 207 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 180-250 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in HEPES, NaCl and Tween®.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile water.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Fibronectin, ACFP Protein

• CIG
• ED-B
• fibronectin 1
• Fibronectin
• FINC
• FN
• FN1
• FNZ
• GFND
• GFND2
• LETS
• MSF
• SMDCF

Background

Fibronectin (FN) is a large, modular glycoprotein that generates a polymeric fibrillar network in the extracellular matrix (ECM), and forms soluble, disulfide-linked dimeric protomers in plasma and other body fluids (1, 2). Fibronectin is a ligand for many molecules, including fibrin, heparin, chondroitin sulfate, collagen/gelatin, and integrins. It is involved in multiple cellular processes such as cell adhesion/migration, blood clotting, morphogenesis, tissue repair, and cell signaling. Fibronectin functions are mediated by the insoluble polymeric fibrillar network. Conversion of soluble Fibronectin to Fibronectin fibrils in the ECM is initiated by binding to cell surface integrins, resulting in exposure of cryptic epitopes necessary for polymerization (1). Fibronectin is made up of three types of homologous structural motifs termed FN type I, type II, and type III repeats (3-5). Alternative splicing generates multiple isoforms of Fibronectin which may have insertions of extra type III domains (EDA and EDB) or alteration of the type III connecting segment (IIICS) (5). Differential splicing within the IIICS domain determines the presence of CS1 and CS2 sequences, and its sensitivity to proteases (6, 7). The tilt angle between type III domains #9 and #10 (which contains an RGD motif) determines integrin binding affinity, suggesting how structural differences between fibrillar and soluble Fibronectin may influence their function (8). From the N-terminus to the furin cleavage site at amino acid 1908, human Fibronectin shares 92% amino acid sequence identity with mouse and rat Fibronectin.

1. Mao, Y. and J.E. Schwarzbauer (2005) Matrix Biol. 24:389.
2. Potts, J.R. and  I.D. Campbell (1996) Matrix Biol. 15:313.
3. Bernard, M.P. et al. (1985) Biochemistry 24:2698.
4. Kornblihtt, A.R. et al. (1983) Proc. Natl. Acad. Sci. USA 80:3218.
5. Kornblihtt, A.R. et al. (1985) EMBO J. 4:1755.
6. Mould, A.P. et al. (1991) J. Biol. Chem. 266:3579.
7. Abe, Y. et al. (2005) Biochem. Biophys. Res. Commun. 338:1640.
8. Altroff, H. et al. (2004) J. Biol. Chem. 279:55995.

Bioinformatics

• Uniprot:
  • CAE45932()