Preparation of Fluorescein Isothiocyanate-Labeled γ-Globulin by Dialysis, Gel Filtration, and Ion-Exchange Chromatography in Combination

Abstract

Dedmon, Robert E. (Presbyterian-St. Luke's Hospital, Chicago, Ill.), Albert W. Holmes, and Friedrich Deinhardt . Preparation of fluorescein isothiocyanate-labeled γ-globulin by dialysis, gel filtration, and ion-exchange chromatography in combination. J. Bacteriol. 89: 734–739. 1965.—Antiviral immune γ-globulins isolated from rabbit and guinea pig sera were labeled through dialysis membranes with fluorescein isothiocyanate and purified in several ways to eliminate nonspecific staining. Gel filtration of the conjugate with Sephadex G-25 coarse beads followed by column fractionation with diethylaminoethyl-Sephadex yielded consistently highly specific staining materials. Fluorescein-protein ratios varied between 1.0 and 4.0. This technique has proved to be simple and reliable, and is less time-consuming than previous techniques.