Flow cytometric measurement of RNA synthesis using bromouridine labelling and bromodeoxyuridine antibodies

Research output: Contribution to journalJournal articlepeer-review

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Flow cytometric measurement of RNA synthesis using bromouridine labelling and bromodeoxyuridine antibodies. / Jensen, P O; Larsen, J; Christiansen, J; Larsen, J K.

In: Cytometry, Vol. 14, No. 4, 1993, p. 455-8.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Jensen, PO, Larsen, J, Christiansen, J & Larsen, JK 1993, 'Flow cytometric measurement of RNA synthesis using bromouridine labelling and bromodeoxyuridine antibodies', Cytometry, vol. 14, no. 4, pp. 455-8. https://doi.org/10.1002/cyto.990140416

APA

Jensen, P. O., Larsen, J., Christiansen, J., & Larsen, J. K. (1993). Flow cytometric measurement of RNA synthesis using bromouridine labelling and bromodeoxyuridine antibodies. Cytometry, 14(4), 455-8. https://doi.org/10.1002/cyto.990140416

Vancouver

Jensen PO, Larsen J, Christiansen J, Larsen JK. Flow cytometric measurement of RNA synthesis using bromouridine labelling and bromodeoxyuridine antibodies. Cytometry. 1993;14(4):455-8. https://doi.org/10.1002/cyto.990140416

Author

Jensen, P O ; Larsen, J ; Christiansen, J ; Larsen, J K. / Flow cytometric measurement of RNA synthesis using bromouridine labelling and bromodeoxyuridine antibodies. In: Cytometry. 1993 ; Vol. 14, No. 4. pp. 455-8.

Bibtex

@article{554cc0e756fe41cbb831b6d762cbd146,
title = "Flow cytometric measurement of RNA synthesis using bromouridine labelling and bromodeoxyuridine antibodies",
abstract = "Nuclear RNA synthesis can be analysed by flow cytometry of cells labelled with 5-bromouridine (BrUrd) and stained with anti-bromodeoxyuridine (BrdUrd) antibody and FITC-conjugated secondary antibody. A panel of 5 different commercially available anti-BrdUrd antibodies was tested on cells of a HL-60 human leukemia cell line, stained as a methanol-fixed nuclear suspension. The BrUrd-induced fluorescence signals were highest with the antibody ABDM (Partec), moderate but reproducible with B-44 (Becton Dickinson), variable or low with BR-3 and IU-4 (Caltag), and not detectable with Bu20a (DAKO). Treatment of BrUrd-labelled nuclei with ribonuclease before staining with antibodies indicated that ABDM and B-44 antibodies specifically recognized BrUrd-substituted RNA, whereas BR-3 and IU-4 antibodies also bound to BrUrd-unlabelled RNA. Combined analysis of BrUrd and DNA contents demonstrated the variation of RNA synthesis during the cell cycle. The BrUrd incorporation was high in the S and G2 phase, variable in G1, and negligible in mitosis. Similar results were obtained using other cell types.",
keywords = "Antibodies, Monoclonal, Bromodeoxyuridine, Flow Cytometry, Fluorescein-5-isothiocyanate, Fluorescent Antibody Technique, Humans, Leukemia, Promyelocytic, Acute, RNA, RNA, Neoplasm, Ribonucleases, Tumor Cells, Cultured, Uridine, Comparative Study, Journal Article, Research Support, Non-U.S. Gov't",
author = "Jensen, {P O} and J Larsen and J Christiansen and Larsen, {J K}",
year = "1993",
doi = "10.1002/cyto.990140416",
language = "English",
volume = "14",
pages = "455--8",
journal = "Cytometry",
issn = "0196-4763",
publisher = "Wiley",
number = "4",

}

RIS

TY - JOUR

T1 - Flow cytometric measurement of RNA synthesis using bromouridine labelling and bromodeoxyuridine antibodies

AU - Jensen, P O

AU - Larsen, J

AU - Christiansen, J

AU - Larsen, J K

PY - 1993

Y1 - 1993

N2 - Nuclear RNA synthesis can be analysed by flow cytometry of cells labelled with 5-bromouridine (BrUrd) and stained with anti-bromodeoxyuridine (BrdUrd) antibody and FITC-conjugated secondary antibody. A panel of 5 different commercially available anti-BrdUrd antibodies was tested on cells of a HL-60 human leukemia cell line, stained as a methanol-fixed nuclear suspension. The BrUrd-induced fluorescence signals were highest with the antibody ABDM (Partec), moderate but reproducible with B-44 (Becton Dickinson), variable or low with BR-3 and IU-4 (Caltag), and not detectable with Bu20a (DAKO). Treatment of BrUrd-labelled nuclei with ribonuclease before staining with antibodies indicated that ABDM and B-44 antibodies specifically recognized BrUrd-substituted RNA, whereas BR-3 and IU-4 antibodies also bound to BrUrd-unlabelled RNA. Combined analysis of BrUrd and DNA contents demonstrated the variation of RNA synthesis during the cell cycle. The BrUrd incorporation was high in the S and G2 phase, variable in G1, and negligible in mitosis. Similar results were obtained using other cell types.

AB - Nuclear RNA synthesis can be analysed by flow cytometry of cells labelled with 5-bromouridine (BrUrd) and stained with anti-bromodeoxyuridine (BrdUrd) antibody and FITC-conjugated secondary antibody. A panel of 5 different commercially available anti-BrdUrd antibodies was tested on cells of a HL-60 human leukemia cell line, stained as a methanol-fixed nuclear suspension. The BrUrd-induced fluorescence signals were highest with the antibody ABDM (Partec), moderate but reproducible with B-44 (Becton Dickinson), variable or low with BR-3 and IU-4 (Caltag), and not detectable with Bu20a (DAKO). Treatment of BrUrd-labelled nuclei with ribonuclease before staining with antibodies indicated that ABDM and B-44 antibodies specifically recognized BrUrd-substituted RNA, whereas BR-3 and IU-4 antibodies also bound to BrUrd-unlabelled RNA. Combined analysis of BrUrd and DNA contents demonstrated the variation of RNA synthesis during the cell cycle. The BrUrd incorporation was high in the S and G2 phase, variable in G1, and negligible in mitosis. Similar results were obtained using other cell types.

KW - Antibodies, Monoclonal

KW - Bromodeoxyuridine

KW - Flow Cytometry

KW - Fluorescein-5-isothiocyanate

KW - Fluorescent Antibody Technique

KW - Humans

KW - Leukemia, Promyelocytic, Acute

KW - RNA

KW - RNA, Neoplasm

KW - Ribonucleases

KW - Tumor Cells, Cultured

KW - Uridine

KW - Comparative Study

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1002/cyto.990140416

DO - 10.1002/cyto.990140416

M3 - Journal article

C2 - 7685681

VL - 14

SP - 455

EP - 458

JO - Cytometry

JF - Cytometry

SN - 0196-4763

IS - 4

ER -

ID: 181874021