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.
CITATION STYLE
Pietrocola, G., Rindi, S., Nobile, G., & Speziale, P. (2017). Purification of human plasma/cellular fibronectin and fibronectin fragments. In Methods in Molecular Biology (Vol. 1627, pp. 309–324). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7113-8_20
Mendeley helps you to discover research relevant for your work.