A gel filtration method for studies on protein iron-binding.

This communication reports the use of a gel filtration technique and radioactive iron for rapid determination of protein-iron binding in complex mixtures. Many methods have been used to investigate the characteristics of iron binding by mammalian serum. Calorimetric techniques have been used to measure the iron-binding capacities of whole seral. Electrophoretic2y3 and precipitation41s methods, using radioactive iron (6aFe), have confirmed the role of a /?-globulin, transferrin, as the primary iron-binding component of serum. Each of these techniques has certain limitations regarding specificity, binding interference, and rapidity of sample processing. Filtration through cross-linked dextran gels separates components of a complex mixture on the basis of molecular dimensions 6 8. The technique has been used for studies of the binding between proteins and small moleculesa*lo.