Human fibronectin has a molecular weight of 450,000 daltons when purified in an intact form from plasma. Fibronectin is a glycoprotein synthesized in the liver for the circulating blood plasma form, and is synthesized by many mesenchymal cells, for the extracellular matrix form. It is composed of two similar, but not identical protein chains, which are linked by two disulfide linkages at the C-terminal end of the chains. The chains are composed of domains which have specific secondary structures linked together by regions which are especially susceptible to proteolytic action. For this reason, detection by immunoblot (western) may show considerable variability in the observed apparent molecular weights, which is predicated on the source of the fibronectin, and to what degree proteolysis has occurred. Bands approximately 225 kDa should be observed after SDS-PAGE when reducing and denaturing conditions are used.
rabbit anti-Fibronectin antibody, FN1, FN, Cold-insoluble globulin, CIG, Anastellin, Ugl-Y1, Ugl-Y2, Ugl-Y3
Fibronectin was purified from Human plasma by binding to a denatured gelatin column followed by elution with high concentrations of arginine. The eluted material was further purified by gel filtration. Immunization occurred after single-band purity was assessed by SDS-PAGE.