Description
Full-length native human plasma fibronectin (~250 kDa subunit, disulfide-linked dimer in solution) purified from screened human plasma. Retains all functional domains — the N-terminal 70 kDa gelatin/collagen-binding region, the central 120 kDa cell-binding module with the RGD recognition motif (FN-III9/10) and the synergy site, the heparin-binding C-terminal 40 kDa region, and the fibrin-binding domains — so the protein is competent for self-polymerisation into fibrils, integrin engagement (a5ß1, aVß3, aVß5), GAG/heparin co-association and matrix cross-talk with collagen, fibrin and other ECM components.
More than 40 published studies have used this product as a coating substrate, micropattern ink, hydrogel additive, antigen, integrin ligand, rescue reagent and patent-protected biofunctional surface component. Two major research lineages — Joachim Rädler / LMU Munich (single-cell micropatterning, 1D migration lanes, two-state patterns, mRNA transfection kinetics) and Adam Cohen / Harvard (optogenetic HEK / cardiac bioelectric tissue) — have built their entire experimental platforms on cat. 663, and their group alumni have carried the protocol to labs across Europe, Asia and North America.
For specific fibronectin sub-domains, see cat. 175 (chymotryptic 120 kDa central cell-attachment fragment) and cat. 815 (40 kDa heparin-binding fragment).
Specifications
Catalog number: 663
Amount: 5 mg
Source: Human plasma (HBsAg / HCV / HIV-1 / HIV-2 negative)
Composition: Full-length plasma fibronectin (250 kDa dimer in solution)
Format: Lyophilized
Functional domains: N-terminal 70 kDa (gelatin / collagen / fibrin binding); central 120 kDa cell-binding (FN-III9/10 RGD + synergy, integrin a5ß1 / aVß3 / aVß5 engagement);
C-terminal 40 kDa heparin / GAG binding; fibrin-binding domains
Reconstitution: Dissolve in 5 ml of warm (37°C) water or PBS. Incubate at 37°C for 3 hours, occasionally inverting the vial. Do not vortex.
Storage: Lyophilized — ambient shipping, -20 °C long-term; reconstituted — short-term at +4 °C, avoid freeze-thaw
APPLICATIONS — overview
- Cell-culture surface coating for adherent mammalian cell lines and primary cells (typical 0.5–50 µg/mL coating concentration).
- Micropatterning (microcontact-printing, plasma-induced patterning, photopatterning) on PDMS, glass, polyacrylamide and silicone substrates for confined cell migration, single-cell trajectory analysis and high-throughput cell behaviour.
- 3D hydrogel functionalisation — polyacrylamide, PEG, silicone, PEGDA and protein-protein cross-linked hydrogels (defined Young's modulus 2–100 kPa).
- Bioelectric / optogenetic tissue engineering — adhesion ligand for spiking HEK monolayers, cardiac and engineered bioelectric tissues.
- mRNA / siRNA transfection kinetics — defined-area FN spots on PEG-passivated micropatterns for single-cell time-lapse transfection assays.
- Integrin-ligand binding assays (QCM, SPR, ELISA) — full-length ligand counterpart to cat. 175 (cell-binding fragment) and the YO Proteins integrin family (cat. 126, 357, 476, 638).
- Endothelial / vascular disease models — rescue substrate for CCM-disrupted, dysfunctional endothelial cells (paired with cat. 175 fragment for domain-mapping).
- Stem-cell expansion and osteogenic / mesenchymal differentiation surfaces.
- Antigen for raising anti-human fibronectin antibodies.
- Biofunctional component in granted patents for cell-array, cell-culture-substrate and tissue-engineering devices.
CITED USE CASES — organised by application theme
The list below summarises 40+ peer-reviewed publications and PhD theses that used YO Proteins cat. 663 (human plasma fibronectin). Within each theme, papers are listed in chronological order. Many of these works state the working concentration directly — typical values are **0.05 mg/mL** for PDMS-stamp microcontact printing (Cohen group), **35–50 µg/mL** for plasma-induced micropatterning (Rädler group), **10 µg/mL** for soft-substrate hydrogel coating (Szeto / Hinz groups), and **1 µg/mL** for sensitive primary-endothelial applications.
### Theme 1 — Single-cell migration and 1D / 2D micropatterning (Rädler LMU lineage)
This is the largest application cluster for cat. 663 — used as the protein ink for plasma-induced micropatterning (PiµP), microcontact printing or photopatterning of fibronectin "lanes" and "two-state" geometries on PEG-passivated surfaces, enabling reproducible single-cell trajectory measurement over thousands of cells.
- Schreiber C. et al. (2016) — Ring-shaped microlanes and chemical barriers as a platform for probing single-cell migration. Scientific Reports 6: 26858.
- Röttgermann P.J.F. (2016) — Single cell motility and apoptosis dynamics on micropatterns. PhD thesis, LMU Munich.
- Reiser A. et al. (2019) — Single-cell mRNA-transfection kinetics on micropatterned arrays. Integrative Biology 11(9): 362–371. doi:10.1093/intbio/zyz030.
- Mitterwallner B.G. et al. (2020) — Non-Markovian data-driven modeling of single-cell motility. Phys Rev E 101: 032408.
- Fink A. (2020) — Cell migration in two-state micropatterns. PhD thesis, LMU Munich.
- Brückner D.B. (2021) — Stochastic dynamics of migrating cells. PhD thesis, LMU Munich.
- Amiri B. et al. (2023) — On multistability and constitutive relations of cell motion on fibronectin lanes. Biophysical Journal.
- Heyn J.C.J. (2024) — Automated time-lapse analysis of cell mechanics in single-cell migration on 1D micropatterns. PhD thesis, LMU Munich.
- Heyn J.C.J. et al. (2025) — Cell-mechanical parameter estimation from 1D cell trajectories using simulation-based inference. PLOS One 20(6): e0310669.
- Jouneau A. et al. (2025) — Inferring three-body interactions in cell migration dynamics. arXiv 2601.05764.
Standard protocol across this body of work: human FN (cat. 663) at **35–50 µg/mL** in PBS, incubated 45–50 min at room temperature on plasma-treated PEG-PLL passivated micropatterns, washed nine times in PBS, stored at +4 °C until cell seeding.
### Theme 2 — Optogenetic bioelectric tissue engineering (Cohen / Harvard lineage)
cat. 663 is the adhesion ligand for confluent monolayers of spiking HEK cells and engineered cardiac tissues used to study propagating bioelectric waves.
- McNamara H.M., Zhang H., Werley C.A., Cohen A.E. (2016) — Optically controlled oscillators in an engineered bioelectric tissue. Phys Rev X 6: 031001.
- Werley C.A., Chien M.-P., Cohen A.E. (2017) — Ultrawidefield microscope for high-speed fluorescence imaging and targeted optogenetic stimulation. Biomedical Optics Express 8(12): 5794–5813.
- McNamara H.M. (2019) — Synthetic physiology: manipulating and measuring biological pattern formation with light. PhD thesis, Harvard University.
Standard protocol: human FN (cat. 663) at **0.05 mg/mL** spotted onto PDMS stamps for 30 min, aspirated, air-dried 10 min before printing onto SU-8 patterned substrates.
### Theme 3 — mRNA / siRNA transfection kinetics on micropatterns
cat. 663-defined FN micro-spots on PEG-passivated surfaces give the "one cell per spot" geometry needed to follow transfection kinetics in thousands of single cells in parallel.
- Leonhardt C.A. (2014) — On quantitative mRNA transfection. PhD thesis, LMU Munich.
- Ferizi M. et al. (2015) — Stability analysis of chemically modified mRNA using micropattern-based single-cell arrays. Lab on a Chip 15: 3561.
- Krzyszton R.S. (2018) — Towards efficient siRNA delivery and gene silencing kinetics on the single-cell level. PhD thesis, LMU Munich.
- Fröhlich F. et al. (2018) — Multi-experiment nonlinear mixed effect modeling of single-cell translation kinetics after transfection. npj Systems Biology and Applications doi:10.1038/s41540-018-0079-7.
- Reiser A. et al. (2019) — Integrative Biology 11(9): 362–371. (cross-listed with Theme 1.)
### Theme 4 — Chip-based single-cell platforms (NK cytotoxicity, cancer, chemotherapy)
- The MainSTREAM Component Platform (2014) — A holistic approach to microfluidic system design (cat. 663 used as the cell-adhesion functionalisation step).
- Chatzopoulou E.I. et al. (2016) — Chip-based platform for dynamic analysis of NK cell cytolysis mediated by a triplebody. Analyst, c5an02585k.
- Chatzopoulou E.I. et al. (2018) — A single-cell micro-trench platform for automatic monitoring of cell division and apoptosis after chemotherapeutic drug administration. Scientific Reports 8: 18042.
Standard protocol: FN (cat. 663) at **35–50 µg/mL** coating PDMS-on-glass chip surfaces for 45–60 min before sterilisation and cell seeding.
### Theme 5 — Mechanobiology — stiffness and topography substrates
- Szeto S.G. et al. (2016) — YAP/TAZ are mechanoregulators of TGF-ß-Smad signaling and renal fibrogenesis. JASN. (cat. 663 at **10 mg/mL** coating 2–100 kPa polyacrylamide/silicone gels for renal fibrosis modelling.)
### Theme 6 — Stem-cell osteogenic / mesenchymal differentiation surfaces
- Padmanabhan H. (2018) — In vitro optimization of osteogenic differentiation outcomes from human mesenchymal stem cells. PhD thesis, University of Queensland.
- Miksiunas R. et al. (2021) — Cardiomyogenic differentiation potential of human dilated myocardium-derived MSCs: HDAC inhibitor SAHA and biomimetic matrices. Int J Mol Sci 22: 12702. (cat. 663 micro-contact-printed as 20 / 45 µm fibronectin lines on PEGMA-hydrogel-coated glass.)
- Velázquez de la Paz M.F. (2022) — Polycaprolactone and poly(glycerol)sebacate polyHIPEs for regeneration of osteochondral defects. PhD thesis, University of Sheffield.
- Hann A.J. (2023) — Development and assessment of in vitro models of osteogenic microstructures. PhD thesis, University of Sheffield.
- Jackson C.E. et al. (2023) — Development of PCL polyHIPE substrates for 3D breast cancer cell culture. Bioengineering 10(5): 522. (cat. 663 as the cell-adhesion functionalisation for PCL-polyHIPE breast-cancer scaffolds.)
### Theme 7 — Vascular / endothelial disease models
- de Kat A.C. et al. (2016) — Early vascular damage in young women with DM-1 and its relation to anti-Müllerian hormone. (cat. 663 at **1 µg/mL** as the gentle coating substrate for primary endothelial expansion on Ibidi grids.)
- (cross-reference: Schwefel et al. 2020 FASEB J — used cat. 175 (120 kDa fragment) head-to-head against full-length cat. 663 in CCM endothelial rescue experiments. See cat. 175 page.)
### Theme 8 — Super-resolution microscopy and reagent characterisation
- Schlichthärle T. (2020) — Expanding the toolbox of DNA-PAINT microscopy: from method development to cellular applications. PhD thesis, LMU Munich. (cat. 663 as a defined cell-adhesion ligand for cellular DNA-PAINT super-resolution imaging.)
### Theme 9 — Cancer cell biology, ECM and migration assays
- Pinon P. et al. (2014) — Talin-bound NPLY motif recruits integrin-signaling adapters... J Cell Biol 205(2): 265–281. (cat. 663 as defined FN substrate in talin-recruitment migration assays.)
- Islam A. et al. (2014) — Acta Biomaterialia (cat. 663 as biofunctional fibronectin surface component.)
- "Cellular self-organization on micro-structured surfaces" (2014) — cat. 663 as the FN ink for micro-structured cell self-organisation assays.
- "Nanoscaled surface patterns influence adhesion" (2014) — cat. 663 as the patterning protein at nanoscale-controlled densities.
- Mat A.M. et al. (2025) — Peripheral tissues of deep-sea mussels exhibit autonomous circadian timing via an atypical mechanism. bioRxiv 681125. (cat. 663 at **1:100 in PBS** as the foot-culture coating substrate.)
- San Felix Garcia-Obregon A. (2023) — Bridging the gap between the mechanical and metabolic activity in cell-ECM interactions. PhD thesis, University of Glasgow. (cat. 663 throughout the thesis as the human-source FN ECM input.)
- Manna A. — PhD thesis, Arizona State University. (cat. 663 in cell-adhesion / mechanobiology studies.)
- Piccitto A. — PhD thesis, Politecnico di Torino. (cat. 663 in biomaterial scaffold functionalisation for diagnostic/therapeutic devices.)
- Sartori S. et al. — Polyurethane scaffolds for the regeneration of the infarcted myocardium. (cat. 663 coupled to polyurethane scaffolds for myocardial regeneration.)
### Theme 10 — Cardiac and developmental biology
- Wagner N., Wagner K.-D. (Eds.) (2022) — Transcriptional regulation of cardiac development and disease. MDPI / IJMS special-issue book. (cat. 663 used in included chapter for FN coating in cardiac-progenitor / cardiomyogenic differentiation.)
- Voelkl I. et al. (2025) — In vitro approaches to study centriole and cilium function in early mouse embryogenesis. (cat. 663 at **10 µg/mL** as the µ-Slide-coating substrate for naive mESC ? EpiLC differentiation.)
### Theme 11 — Vascular smooth muscle / atherosclerosis
- Ferris G.R. (2025) — Dissecting the regulation of vascular matrix component production and the contribution it plays in endothelial erosion. PhD thesis, Manchester Metropolitan University. (cat. 663 throughout PhD thesis on vascular ECM biology.)
### Theme 12 — Granted patent
- EP 3 416 695 B1 (granted 2018) — uses **0.1 mg/mL human fibronectin (Yo Proteins AB)** in 0.1 M PBS pH 8.0 mixed with HiLyte-Fluor-488-labelled bovine plasma FN as the patterning "ink" in a granted European patent on cell-array / cell-culture-substrate devices. This is a strong commercial-validation signal — cat. 663 is the specifically named patent-pedigree FN reagent.
### Theme 13 — Other documented uses
- London PhD thesis (U591916) — Anti-basal ganglia antibodies in movement disorders. (cat. 663 used as the FN reference in immunoassays.)
FAQ: Handling and Storage of Fibronectin
How do you dissolve lyophilized fibronectin?
Add 5 ml of warm PBS or water (37°C) to the lyophilized vial. Incubate at 37°C for 3 hours, inverting the vial periodically. Do not shake or vortex — shear force causes aggregation.
The reconstituted solution contains 5 mg soluble fibronectin at 1 mg/ml (absorbance A280 = 1.3) plus a small insoluble aggregate fraction. The vial is supplied slightly overfilled to account for this. Filter slowly through a prewetted 0.2 µm membrane filter.
Can you freeze reconstituted fibronectin?
No. Despite advice to the contrary that circulates in the literature, we strongly recommend against freezing reconstituted fibronectin. Fibronectin is a large, flexible glycoprotein that aggregates irreversibly after freeze-thaw cycling. Aggregated fibronectin coats unevenly and can compromise cell attachment assays.
Lyophilized fibronectin is stable at -20°C for up to 2 years and is reasonably stable at ambient temperature during shipping. Once reconstituted and sterile filtered, store at +4°C and use within 1 month. Always reconstitute freshly where possible.
What concentration should I use for cell culture coating?
For most standard cell culture applications, 1–10 µg/cm² is the typical coating range. Common working protocols:
- General cell attachment: 1–5 µg/cm² in PBS, incubate 1 hour at room temperature or overnight at +4°C
- Serum-free and defined media culture: 5–10 µg/cm²
- Micropatterning and soft lithography: concentration depends on stamp geometry — 50 µg/ml solution is standard for PDMS microcontact printing
- iPSC and stem cell culture: 5 µg/cm² is widely reported in the literature
