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Fibrosis pp 309-324 | Cite as

Purification of Human Plasma/Cellular Fibronectin and Fibronectin Fragments

  • Giampiero Pietrocola
  • Simonetta Rindi
  • Giulia Nobile
  • Pietro Speziale
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1627)

Abstract

A method is described for the purification of plasma fibronectins based on a combination of gelatin- and arginine-Sepharose chromatography steps. Cellular fibronectin can be purified from an osteosarcoma fibroblast cell line by affinity chromatography using a monoclonal antibody anti-fibronectin as ligand. Furthermore, we also provide a protocol for the purification of fibronectin domains obtained by fractionation of thermolysin-digested plasma fibronectin on ion-exchange/gel filtration chromatography columns. Assessment of the fibronectin purity is performed by SDS-PAGE, while the ligand binding activities of specific fibronectin domains are determined by ELISA.

Key words

Fibronectin Fibronectin purification Affinity chromatography Fibronectin domain Ion-exchange chromatography Gel filtration chromatography SDS-PAGE ELISA 

Notes

Acknowledgments

This work was supported in part by Fondazione CARIPLO Grant Vaccines 2009-3546 to P.S.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Giampiero Pietrocola
    • 1
  • Simonetta Rindi
    • 1
  • Giulia Nobile
    • 1
  • Pietro Speziale
    • 1
  1. 1.Unit of Biochemistry, Department of Molecular MedicineUniversity of PaviaPaviaItaly

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