Human fibronectin 120 kDa cell-attachment fragment, 0.5 mg — the chymotryptic fragment carrying the central RGD cell-binding domain, purified from plasma fibronectin. It presents the integrin-engagement function of fibronectin in isolation, without the matrix-polymerisation, heparin/GAG-binding and collagen-association activities of the full-length molecule, making it the reagent of choice when a phenotype must be attributed to integrin engagement per se. For full-length human plasma fibronectin (250 kDa dimer) see cat. 663; for the heparin-binding fragment see cat. 815.
SPECIFICATIONS
Catalogue number: 175 | Source: Human plasma (negative for HBsAg, HCV, HIV-1 and HIV-2)
Purity: One major band at 120 kDa on SDS-PAGE; no bands above 120 kDa
Purification: Chymotryptic digest of purified human plasma fibronectin; affinity chromatography on gelatin-agarose, then heparin-agarose, followed by gel filtration
Functional sites: Central RGD cell-attachment site; integrin a5b1 binding (FN-III9/10 RGD + synergy site); integrin aVb3 binding
Format: Lyophilised from sodium citrate buffer, pH 7.4 | Package size: 0.5 mg
STORAGE / RECONSTITUTION
Add 0.5 mL of sterile pure water to the vial, incubate 15 min at 37 °C and stir gently; do not vortex. Ships at ambient temperature. Store lyophilised protein up to 1 year at −20 °C or lower; store reconstituted protein up to 6 months at −70 °C. Avoid repeated freeze-thaw.
APPLICATIONS
- Cell-attachment and spreading assays — rapid, efficient adhesion of integrin-expressing cell lines and primary cells; recommended coating concentration 5 µg/cm².
- Integrin-signalling assays — defined ligand for outside-in signalling through a5b1 and aVb3 (FAK activation, paxillin / vinculin recruitment, Rho-family GTPase activity). Purified integrins are available as cat. 638 (a5b1), cat. 357 (aVb3) and cat. 476 (aVb5).
- Rescue / loss-of-function assays — isolates the integrin-engagement contribution of fibronectin. Use as a coating substrate (typically 5 µg/cm²) or supplement to growth medium (typically 32 µg/mL).
- Endothelial-cell phenotype rescue — restores spheroid organisation and cytoskeletal integrity in fibronectin-deficient or CCM-disrupted endothelial cells.
- Biomaterial functionalisation — RGD-presenting ligand for coupling to synthetic scaffolds, hydrogels and microfluidic chip surfaces.
- Competitor / inhibitor reagent in cell-adhesion blocking studies, alongside anti-integrin antibodies or RGD peptides.
- Antigen for raising anti-fibronectin antibodies against the central cell-binding domain.
APPLICATION HIGHLIGHTS — HIGH-VALUE RESEARCH NICHES
- Integrin biology — dissecting RGD-dependent adhesion from fibrillogenesis and GAG binding.
- Vascular malformation research — CCM1/2/3-deficient endothelial phenotype rescue.
- Cancer cell migration and invasion — defined adhesion substrate for motility assays.
- Stem-cell and iPSC culture — xeno-defined RGD coating alternative to full-length matrices.
- Biomaterials and organ-on-chip — minimal adhesive ligand for surface functionalisation.
REFERENCES
Schwefel K. et al. (2020) Fibronectin rescues aberrant phenotype of endothelial cells lacking either CCM1, CCM2 or CCM3. The FASEB Journal 34(7): 9018–9033. doi:10.1096/fj.201902888R.
