Abstract
This technique is used to visualize sites of active transcription in a permeabilized cell and does not require radiolabeled molecules (e.g., [3H] Uridine). Nonradioactive ribonucleic acid (RNA) precursors (e.g., 5-bromouridine-5′-triphosphate [BrUTP]) are used and can be detected by using fluorescently labeled antibodies. Procedures for BrUTP of labeling transcription sites require manipulations that are best applied to adherent cells but can be applied, with difficulty, to cell cultures in suspension. The protocol described below is for adherent cells grown on cover slips.
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References
Wansink, D. G., Motley, A. M., van Driel, R., and de Jong, L. (1994) Fluorescent labeling of nascent RNA in the cell nucleus using 5-bromouridine 5′-triphosphate. in Cell Biology—A Laboratory Handbook (J. Celis, ed.), Academic Press, San Diego, CA, pp. 368–374.
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© 2004 Humana Press Inc.
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Javed, A. et al. (2004). In Situ Immunofluorescence Analysis. In: Giordano, A., Romano, G. (eds) Cell Cycle Control and Dysregulation Protocols. Methods in Molecular Biology™, vol 285. Humana Press. https://doi.org/10.1385/1-59259-822-6:029
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DOI: https://doi.org/10.1385/1-59259-822-6:029
Publisher Name: Humana Press
Print ISBN: 978-0-89603-949-0
Online ISBN: 978-1-59259-822-9
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