{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T23:44:06Z","timestamp":1790984646668,"version":"4.1.0"},"reference-count":27,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2005,3,8]],"date-time":"2005-03-08T00:00:00Z","timestamp":1110240000000},"content-version":"vor","delay-in-days":3994,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry"],"published-print":{"date-parts":[[1994,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The currently available flow cytometric stains for cellular glutathione were evaluated, examining the labelling of both human and rodent cell lines under various conditions of concentration, time, and temperature. Procedures were used that depleted, glutathione (GSH) while having a minimal effect on other cellular sulphydryls in order to estimate linearity and the extent of background staining. As previously reported, monochlorobimane was highly specific for GSH in rodent cells but failed to label human cells adequately because of its low affinity for human glutathione S\u2010transferases. Higher concentrations of monochlorobimane achieved more complete labelling of, the human cellular GSH pool but gave increased background fluorescence due to non\u2010GSH binding. The analogue monobromobimane, which binds nonenzymatically to sulphydryls, reacted, more readily with GSH than with protein sulphydryls and, provided that stain concentration and incubation time were controlled, gave reproducible staining of human cells with 20% of total fluorescence due to background staining. Of the currently available stains for measuring GSH in human cells, monobromobimane is the agent of choice. Mercury orange also binds more readily to GSH than to protein, giving a degree of specificity, and it has the additional advantage of being excited at 488 nm. However, the reproducibility of staining with mercury orange was less consistent than that using monobromobimane, and a higher background fluorescence was seen. Two additional stains, O\u2010phthaldialdehyde and chloromethyl fluorescein, could also be used to label cellular GSH, but both gave an unacceptably high level of background staining. It is recommended that flow cytometric GSH assays should routinely include a sample of cells that have been depleted of GSH in order to determine the extent of background labelling. \u00a9 1994 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/cyto.990150411","type":"journal-article","created":{"date-parts":[[2005,6,30]],"date-time":"2005-06-30T19:55:53Z","timestamp":1120161353000},"page":"349-358","source":"Crossref","is-referenced-by-count":203,"title":["Evaluation of methods for measuring cellular glutathione content using flow cytometry"],"prefix":"10.1002","volume":"15","author":[{"given":"David W.","family":"Hedley","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sue","family":"Chow","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2005,6,23]]},"reference":[{"key":"e_1_2_1_2_2","first-page":"4224","article-title":"Glutathione metabolism as a determinant of therapeutic efficacy: A review","volume":"44","author":"Arrick BA","year":"1984","journal-title":"Cancer Res"},{"key":"e_1_2_1_3_2","doi-asserted-by":"publisher","DOI":"10.1177\/23.10.53246"},{"key":"e_1_2_1_4_2","doi-asserted-by":"publisher","DOI":"10.1002\/ar.1091100208"},{"key":"e_1_2_1_5_2","doi-asserted-by":"publisher","DOI":"10.1097\/00008390-199201000-00003"},{"key":"e_1_2_1_6_2","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.1988.6.4.709"},{"key":"e_1_2_1_7_2","first-page":"1696","article-title":"Differential specificity of monochlorobimane for isozymes of human and rodent glutathione S\u2010transferases","volume":"51","author":"Cook JA","year":"1991","journal-title":"Cancer Res"},{"key":"e_1_2_1_8_2","first-page":"4287","article-title":"Cellular glutathione and thiol measurements from surgically resected human lung tumor and normal lung tissue","volume":"51","author":"Cook JA","year":"1991","journal-title":"Cancer Res"},{"key":"e_1_2_1_9_2","doi-asserted-by":"publisher","DOI":"10.2307\/3576093"},{"key":"e_1_2_1_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(86)90061-8"},{"key":"e_1_2_1_11_2","unstructured":"HauglandRP:Handbook of Fluorescent Probes and Research Chemicals. 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