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
Hauptverfasser: Bakker, P. J. M., Stap, J., Tukker, C. J., van Oven, C. H., Veenhof, C. H. N., Aten, J.
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container_end_page 372
container_issue 4
container_start_page 366
container_title Cytometry (New York, N.Y.)
container_volume 12
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.</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 &amp; 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. 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source MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection
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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