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Labeling of small molecules with fluorescein

Labeling of small molecules with fluorescein

Analytical Biochemistry 89(1): 304-305

The finding that fluorescein passes through a cell junction of low electrical resistance, has led to the synthesis of other fluorescent molecules for probing junctional permeability. These new probes combined some of the desirable features of fluorescein, namely a high fluorescence yield, excitation by visible light, low toxicity toward cells and lack of nonjunctional membrane transport, with some nonfluorescent parameters, e.g., size and charge. Thus fluorescein isothiocyanate (FITC) which has been widely used in immunochemistry to label proteins was coupled to some smaller substances of well-defined structure such as amino acids. Although the actual coupling of FITC to small substrates presents no problem, the separation of the coupled product from the hydrolysed dye can be a major difficulty. Details of a simple chromatographic method for carrying out this separation are described, and the method is illustrated with insulin A chain.

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Accession: 005787059

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PMID: 707804

DOI: 10.1016/0003-2697(78)90755-8

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