The Staining Properties of Human Serum Proteins Conjugated with Purified Fluorescein Isothiocyanate

Open Access
Publisher Website
Google Scholar
Abstract
Summary: Electrophoretic analysis has shown that six preparations of fluorescein isothiocyanate (FITC) from one source contain degradation products which exhibit essentially similar fluorescence properties and a similar capacity to couple with serum protein, but which confer upon the conjugate the propensity for nonspecific staining. It is recommended that only undegraded FITC (e.g., BBL) which manifests superior specificity and higher fluorescence intensity, be used for the conjugation of specific immune γ-globulins, and that the pH of the conjugation system should not exceed 8.8. Purified FITC is now commerically available or may be easily obtained by continuous flow electrophoresis. It is emphasized, however, that the degradation products of FITC constitute only one of a number of factors contributing to nonspecific fluorescent staining. Two other such factors, relating to the overlabeling of the γ-globulin and to the other serum proteins, are discussed.
Keywords

This publication has 0 references indexed in Scilit: