caa a22 4500 469030860 CHVBK 20180323132743.0 cr unu---uuuuu 170328e19920501xx s 000 0 eng 10.1007/BF01046839 doi (NATIONALLICENCE)springer-10.1007/BF01046839 DNA double labelling with IdUrd and CldUrd for spatial and temporal analysis of cell proliferation and DNA replication [Elektronische Daten] [J. Aten, P. Bakker, J. Stap, G. Boschman, C. Veenhof] Summary: A procedure was developed that very effectively distinguishes between IdUrd and CldUrd incorporated in the DNA of cell nuclei and chromosomes. For double staining we used the, rat anti-BrdUrd monoclonal antibody from Sera-lab that binds specifically to CldUrd and BrdUrd but not to IdUrd, in combination with the mouse anti-BrdUrd monoclonal antibody from Becton Dickinson. This antibody binds to all three halogenated deoxyuridines, but when the nuclei are washed in TRIS buffer with a high salt concentration the antibodies linked to CldUrd-labelled DNA are removed. When analysing the effect of the deoxyuridines on the cell cycle we found that the growth kinetics of Chinese hamster cells were not changed by adding IdUrd or CldUrd for 30 min at a concentration of 10 μm, whereas adequate double labelling required only 2 min pulses. The effectiveness of the technique was demonstrated in two model experiments. The first test concerned the assessment of cell recruitment in the central areas of slow-growing clones, after addition of fresh medium. The second experiment focussed on the spatial resolution of the method. Double-labelled metaphase chromosomes showed interspersed green and red replication bands with a spacing corresponding with medium resolution Giemsa banding patterns. Chapman & Hall, 1992 Aten J. Laboratory for Radiobiology, Department of Internal Medicine, University of Amsterdam, Academic Medical Centre, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands aut Bakker P. Division of Medical Oncology, Department of Internal Medicine, University of Amsterdam Academic Medical Centre, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands aut Stap J. Laboratory for Radiobiology, Department of Internal Medicine, University of Amsterdam, Academic Medical Centre, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands aut Boschman G. Laboratory for Radiobiology, Department of Internal Medicine, University of Amsterdam, Academic Medical Centre, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands aut Veenhof C. Division of Medical Oncology, Department of Internal Medicine, University of Amsterdam Academic Medical Centre, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands aut The Histochemical Journal Kluwer Academic Publishers 24/5(1992-05-01), 251-259 0018-2214 24:5<251 1992 24 10735 https://doi.org/10.1007/BF01046839 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF01046839 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Aten J. Laboratory for Radiobiology, Department of Internal Medicine, University of Amsterdam, Academic Medical Centre, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands aut NATIONALLICENCE 700 1- Bakker P. Division of Medical Oncology, Department of Internal Medicine, University of Amsterdam Academic Medical Centre, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands aut NATIONALLICENCE 700 1- Stap J. Laboratory for Radiobiology, Department of Internal Medicine, University of Amsterdam, Academic Medical Centre, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands aut NATIONALLICENCE 700 1- Boschman G. Laboratory for Radiobiology, Department of Internal Medicine, University of Amsterdam, Academic Medical Centre, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands aut NATIONALLICENCE 700 1- Veenhof C. Division of Medical Oncology, Department of Internal Medicine, University of Amsterdam Academic Medical Centre, Meibergdreef 9, 1105 AZ, Amsterdam, The Netherlands aut NATIONALLICENCE 773 0- The Histochemical Journal Kluwer Academic Publishers 24/5(1992-05-01), 251-259 0018-2214 24:5<251 1992 24 10735 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer