Purification of Human Plasma/Cellular Fibronectin and Fibronectin Fragments
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 wordsFibronectin Fibronectin purification Affinity chromatography Fibronectin domain Ion-exchange chromatography Gel filtration chromatography SDS-PAGE ELISA
This work was supported in part by Fondazione CARIPLO Grant Vaccines 2009-3546 to P.S.
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