{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T15:14:38Z","timestamp":1764688478425},"reference-count":16,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2005,3,8]],"date-time":"2005-03-08T00:00:00Z","timestamp":1110240000000},"content-version":"vor","delay-in-days":7067,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry"],"published-print":{"date-parts":[[1985,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>This report describes an improved immunochemical procedure for staining cells in suspension for amount of incorporated bromodeoxyuridine (BrdUrd) and total DNA. In this procedure, cellular DNA is partially denatured by extracting the cells with 0. 1 M HCl and then heating them to 80\u00b0C in a 50% formamide solution. The cells are then immuno\u2010fluorescently stained using a monoclonal antibody against BrdUrd in single\u2010strand DNA (ssDNA) and counterstained for DNA content with propidium iodide (PI), a dye that fluoresces preferentially when bound to double\u2010strand DNA (dsDNA). We show that the relative amounts of immunofluorescently stained BrdUrd in ssDNA and PI in dsDNA can be altered reciprocally by changing the formamide concentration, denaturation time, and denaturation temperature. We show that this new immunochemical staining procedure allows more complete DNA denaturation so that fivefold lower levels of BrdUrd incorporation can be quantified. In addition, we show that the BrdUrd\u2010linked immunofluorescence achieved using the new denaturation procedure is more linearly related to cellular BrdUrd content than that achieved after acid DNA denaturation. However, cell loss is sufficiently severe with the thermal denaturation procedure that it may not be applicable to all cell types.<\/jats:p>","DOI":"10.1002\/cyto.990060606","type":"journal-article","created":{"date-parts":[[2005,8,8]],"date-time":"2005-08-08T14:56:33Z","timestamp":1123512993000},"page":"521-530","source":"Crossref","is-referenced-by-count":123,"title":["Cytochemistry for bromodeoxyuridine\/DNA analysis: Stoichiometry and sensitivity"],"prefix":"10.1002","volume":"6","author":[{"given":"F.","family":"Dolbeare","sequence":"first","affiliation":[]},{"given":"W.","family":"Beisker","sequence":"additional","affiliation":[]},{"given":"M. G.","family":"Pallavicini","sequence":"additional","affiliation":[]},{"given":"M.","family":"Vanderlaan","sequence":"additional","affiliation":[]},{"given":"J. W.","family":"Gray","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2005,6,16]]},"reference":[{"key":"e_1_2_1_2_2","first-page":"427","article-title":"Cell\u2010cycle analysis using a monoclonal antibody to BrdU","volume":"17","author":"Dean PN","year":"1984","journal-title":"Cell Tissue Kinet"},{"key":"e_1_2_1_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.80.18.5573"},{"key":"e_1_2_1_4_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.7123245"},{"key":"e_1_2_1_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/0163-7258(83)90003-7"},{"key":"e_1_2_1_6_2","volume-title":"Techniques in Cell Cycle Analysis","author":"Gray JW","year":"1984"},{"key":"e_1_2_1_7_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja01462a027"},{"key":"e_1_2_1_8_2","first-page":"539","article-title":"Factors influencing the quantitative estimation of the in vivo survival of cells from solid tumors","volume":"39","author":"Kallman RF","year":"1967","journal-title":"J Natl Cancer Inst"},{"key":"e_1_2_1_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/0006-3002(61)91013-7"},{"key":"e_1_2_1_10_2","doi-asserted-by":"crossref","unstructured":"MoranR DarzynkiewiczZ Staiano\u2010coicoLMelamedMR: Detection of BrdUrd incorporation by monoclonal antibodies: Role of the denaturation step.J Histochem Cytochem(in press) 1985.","DOI":"10.1177\/33.8.3860561"},{"key":"e_1_2_1_11_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.990020203"},{"key":"e_1_2_1_12_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.990060608"},{"key":"e_1_2_1_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.990060616"},{"key":"e_1_2_1_14_2","doi-asserted-by":"publisher","DOI":"10.2172\/5842852"},{"key":"e_1_2_1_15_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM198404123101518"},{"key":"e_1_2_1_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0769-2625(83)80030-0"},{"key":"e_1_2_1_17_2","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1002\/cyto.990060603","article-title":"Characterization of monoclonal antibodies to bromodeoxyuridine","volume":"6","author":"Vanderlaan M","journal-title":"Cytometry"}],"container-title":["Cytometry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fcyto.990060606","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cyto.990060606","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,13]],"date-time":"2023-09-13T14:55:29Z","timestamp":1694616929000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/cyto.990060606"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1985,11]]},"references-count":16,"journal-issue":{"issue":"6","published-print":{"date-parts":[[1985,11]]}},"alternative-id":["10.1002\/cyto.990060606"],"URL":"https:\/\/doi.org\/10.1002\/cyto.990060606","archive":["Portico"],"relation":{},"ISSN":["0196-4763","1097-0320"],"issn-type":[{"value":"0196-4763","type":"print"},{"value":"1097-0320","type":"electronic"}],"subject":[],"published":{"date-parts":[[1985,11]]}}}