An indirect immunofluorescence double staining procedure for the simultaneous flow cytometric measurement of iodo‐ and chlorodeoxyuridine incorporated into DNA
In this paper we describe an indirect fluorescence double staining procedure for the simultaneous detection of IdUrd and CldUrd in the same cell nucleus. Two commercially available antibodies were selected for this purpose. A rat antiBrdUrd monoclonal antibody from Seralab was found to bind specific...
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Veröffentlicht in: | Cytometry (New York, N.Y.) N.Y.), 1991, Vol.12 (4), p.366-372 |
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creator | Bakker, P. J. M. Stap, J. Tukker, C. J. van Oven, C. H. Veenhof, C. H. N. Aten, J. |
description | In this paper we describe an indirect fluorescence double staining procedure for the simultaneous detection of IdUrd and CldUrd in the same cell nucleus. Two commercially available antibodies were selected for this purpose. A rat antiBrdUrd monoclonal antibody from Seralab was found to bind specifically to CldUrd and BrdUrd. A mouse monoclonal anti‐BrdUrd antibody from Becton Dickinson used in a 1:2 dilution binds to all halogenated deoxyuridines but, when the cells were extensively washed with Tris buffer with a high salt concentration, almost no binding to CldUrd was observed. An immunofluorescence procedure was developed, based on these primary antibodies, raised in different species (rat and mouse), in combination with highly purified second antibodies: FITC conjugated goat antirat and Texas‐Red conjugated goat antimouse. |
doi_str_mv | 10.1002/cyto.990120412 |
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J. M. ; Stap, J. ; Tukker, C. J. ; van Oven, C. H. ; Veenhof, C. H. N. ; Aten, J.</creator><creatorcontrib>Bakker, P. J. M. ; Stap, J. ; Tukker, C. J. ; van Oven, C. H. ; Veenhof, C. H. N. ; Aten, J.</creatorcontrib><description>In this paper we describe an indirect fluorescence double staining procedure for the simultaneous detection of IdUrd and CldUrd in the same cell nucleus. Two commercially available antibodies were selected for this purpose. A rat antiBrdUrd monoclonal antibody from Seralab was found to bind specifically to CldUrd and BrdUrd. A mouse monoclonal anti‐BrdUrd antibody from Becton Dickinson used in a 1:2 dilution binds to all halogenated deoxyuridines but, when the cells were extensively washed with Tris buffer with a high salt concentration, almost no binding to CldUrd was observed. An immunofluorescence procedure was developed, based on these primary antibodies, raised in different species (rat and mouse), in combination with highly purified second antibodies: FITC conjugated goat antirat and Texas‐Red conjugated goat antimouse.</description><identifier>ISSN: 0196-4763</identifier><identifier>EISSN: 1097-0320</identifier><identifier>DOI: 10.1002/cyto.990120412</identifier><identifier>PMID: 2065560</identifier><identifier>CODEN: CYTODQ</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Animals ; Biological and medical sciences ; cell kinetics ; Cells, Cultured ; CldUrd ; Cricetinae ; Cricetulus ; Deoxyuridine - analogs & derivatives ; Deoxyuridine - analysis ; Diverse techniques ; DNA - chemistry ; double staining procedure ; flow cytometry ; Flow Cytometry - methods ; Fluorescent Antibody Technique ; Fundamental and applied biological sciences. Psychology ; Idoxuridine - analysis ; IdUrd ; Immunochemistry ; Mice ; Molecular and cellular biology</subject><ispartof>Cytometry (New York, N.Y.), 1991, Vol.12 (4), p.366-372</ispartof><rights>Copyright © 1991 Wiley‐Liss, Inc.</rights><rights>1992 INIST-CNRS</rights><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3242-c8a5540897e9dd67c5bcce772557271d830fa072c0a78a61390e69b435e4cf183</citedby><cites>FETCH-LOGICAL-c3242-c8a5540897e9dd67c5bcce772557271d830fa072c0a78a61390e69b435e4cf183</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,4022,27922,27923,27924</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=5312988$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2065560$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bakker, P. J. M.</creatorcontrib><creatorcontrib>Stap, J.</creatorcontrib><creatorcontrib>Tukker, C. J.</creatorcontrib><creatorcontrib>van Oven, C. H.</creatorcontrib><creatorcontrib>Veenhof, C. H. N.</creatorcontrib><creatorcontrib>Aten, J.</creatorcontrib><title>An indirect immunofluorescence double staining procedure for the simultaneous flow cytometric measurement of iodo‐ and chlorodeoxyuridine incorporated into DNA</title><title>Cytometry (New York, N.Y.)</title><addtitle>Cytometry</addtitle><description>In this paper we describe an indirect fluorescence double staining procedure for the simultaneous detection of IdUrd and CldUrd in the same cell nucleus. Two commercially available antibodies were selected for this purpose. A rat antiBrdUrd monoclonal antibody from Seralab was found to bind specifically to CldUrd and BrdUrd. A mouse monoclonal anti‐BrdUrd antibody from Becton Dickinson used in a 1:2 dilution binds to all halogenated deoxyuridines but, when the cells were extensively washed with Tris buffer with a high salt concentration, almost no binding to CldUrd was observed. An immunofluorescence procedure was developed, based on these primary antibodies, raised in different species (rat and mouse), in combination with highly purified second antibodies: FITC conjugated goat antirat and Texas‐Red conjugated goat antimouse.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>cell kinetics</subject><subject>Cells, Cultured</subject><subject>CldUrd</subject><subject>Cricetinae</subject><subject>Cricetulus</subject><subject>Deoxyuridine - analogs & derivatives</subject><subject>Deoxyuridine - analysis</subject><subject>Diverse techniques</subject><subject>DNA - chemistry</subject><subject>double staining procedure</subject><subject>flow cytometry</subject><subject>Flow Cytometry - methods</subject><subject>Fluorescent Antibody Technique</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Idoxuridine - analysis</subject><subject>IdUrd</subject><subject>Immunochemistry</subject><subject>Mice</subject><subject>Molecular and cellular biology</subject><issn>0196-4763</issn><issn>1097-0320</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1991</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkbGOEzEQhi0EOsJBS4fkAtEljO1de11GgQOkE9ccBdXKsWc5I68dbK-OdPcIvAKvxpOwUaJQUo2s__PMP_MT8pLBigHwt3Zf00prYBwaxh-RBQOtliA4PCYLYFouGyXFU_KslO8AoGUjLsgFB9m2Ehbk9zpSH53PaCv14zjFNIQpZSwWo0Xq0rQNSEs1Pvr4je5ysuimjHRImda7WfLjFKqJmKZCh5Du6cHSiDV7S0c0ZYZHjJWmgfrk0p-HX9RER-1dSDk5TD_3U_bOR5yN2JR3KZuKbn7URN99Xj8nTwYTCr441Uvy5er97ebj8vrmw6fN-nppBW_40nambRvotELtnFS23VqLSvG2VVwx1wkYDChuwajOSCY0oNTbRrTY2IF14pK8OfadV_wxYan96OcjhHBcre9AStE1YgZXR9DmVErGod9lP5q87xn0h0z6wwH6cybzh1enztN2RHfGTyHM-uuTboo1YcgmWl_OWCsY193BoD5i9z7g_j9D-83X25t_Fv4ChsSrFg</recordid><startdate>1991</startdate><enddate>1991</enddate><creator>Bakker, P. J. M.</creator><creator>Stap, J.</creator><creator>Tukker, C. J.</creator><creator>van Oven, C. H.</creator><creator>Veenhof, C. H. N.</creator><creator>Aten, J.</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley-Liss</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>1991</creationdate><title>An indirect immunofluorescence double staining procedure for the simultaneous flow cytometric measurement of iodo‐ and chlorodeoxyuridine incorporated into DNA</title><author>Bakker, P. J. M. ; Stap, J. ; Tukker, C. J. ; van Oven, C. H. ; Veenhof, C. H. N. ; Aten, J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3242-c8a5540897e9dd67c5bcce772557271d830fa072c0a78a61390e69b435e4cf183</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1991</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>cell kinetics</topic><topic>Cells, Cultured</topic><topic>CldUrd</topic><topic>Cricetinae</topic><topic>Cricetulus</topic><topic>Deoxyuridine - analogs & derivatives</topic><topic>Deoxyuridine - analysis</topic><topic>Diverse techniques</topic><topic>DNA - chemistry</topic><topic>double staining procedure</topic><topic>flow cytometry</topic><topic>Flow Cytometry - methods</topic><topic>Fluorescent Antibody Technique</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Idoxuridine - analysis</topic><topic>IdUrd</topic><topic>Immunochemistry</topic><topic>Mice</topic><topic>Molecular and cellular biology</topic><toplevel>online_resources</toplevel><creatorcontrib>Bakker, P. J. M.</creatorcontrib><creatorcontrib>Stap, J.</creatorcontrib><creatorcontrib>Tukker, C. J.</creatorcontrib><creatorcontrib>van Oven, C. H.</creatorcontrib><creatorcontrib>Veenhof, C. H. N.</creatorcontrib><creatorcontrib>Aten, J.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Cytometry (New York, N.Y.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bakker, P. J. 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A mouse monoclonal anti‐BrdUrd antibody from Becton Dickinson used in a 1:2 dilution binds to all halogenated deoxyuridines but, when the cells were extensively washed with Tris buffer with a high salt concentration, almost no binding to CldUrd was observed. An immunofluorescence procedure was developed, based on these primary antibodies, raised in different species (rat and mouse), in combination with highly purified second antibodies: FITC conjugated goat antirat and Texas‐Red conjugated goat antimouse.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>2065560</pmid><doi>10.1002/cyto.990120412</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Animals Biological and medical sciences cell kinetics Cells, Cultured CldUrd Cricetinae Cricetulus Deoxyuridine - analogs & derivatives Deoxyuridine - analysis Diverse techniques DNA - chemistry double staining procedure flow cytometry Flow Cytometry - methods Fluorescent Antibody Technique Fundamental and applied biological sciences. Psychology Idoxuridine - analysis IdUrd Immunochemistry Mice Molecular and cellular biology |
title | An indirect immunofluorescence double staining procedure for the simultaneous flow cytometric measurement of iodo‐ and chlorodeoxyuridine incorporated into DNA |
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