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Methods Enzymol. 1999;299:247-58.

Flow cytometric determination of cellular thiols.

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University of California, Berkeley 94720-3200, USA.


Several biochemical techniques are based on chromatography or electrophoresis for the determination of thiols from biological samples. These techniques are indispensable for the accurate and sensitive detection of specific thiols. Flow cytometric determination of cellular thiols is a powerful technique that is perhaps best suited for clinical application, particularly for cells in blood or other body fluids. Information can be obtained from a small sample amount with a relatively little and quick sample treatment. This technique offers an unique advantage to study the thiol status of a subset of cells because data are collected from individual cells. Multiparameter flow cytometry allows the study of different subsets of immunotyped cells. A major drawback of the flow cytometric method is the lack of specificity for the determination of distinct thiols. The reaction between MBB and thiols is not specific for any particular intracellular thiol, although almost all of the entire thiol-reacted bimane emission is specific for thiols in general. This limitation can be partly overcome by the treatment of cells with known thiol regulatory agents as described in the section on the differential assessment of cellular thiols.

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