A Kinetic Study on the Labeling of Serum Globulin with Fluorescein Isothiocyanate by Means of the Gel Filtration Technique

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Abstract
Summary: It was found that Sephadex gel filtration could be used for the quantitative analysis of reactants by rapid separation. Using simultaneous gel filtration of samples in a number of sister columns association constants and the number of sites were obtained for the kinetics of the labeling process of globulins with fluorescein. Reagents for protein precipitation were used to remove superficially adsorbed dye from fluorescent atibody. The results of such precipitation showed that the gel-filtered material contained smaller amounts of adsorbed dye than dialyzed material. The patterns of DEAE-cellulose and DEAE-Sephadex column chromatography were compared by the use of fluorescent antisera. It was shown that the complex of papain and fluorescent antibody could be separated from free papain satisfactorily on a Sephadex G-75 column.

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