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Quantitative imaging assay for NF-kappa B nuclear translocation in primary human macrophages

Noursadeghi, M; Tsang, J; Haustein, T; Miller, RF; Chain, BM; Katz, DR; (2008) Quantitative imaging assay for NF-kappa B nuclear translocation in primary human macrophages. Journal of Immunological Methods , 329 (1-2) 194 - 200. 10.1016/j.jim.2007.10.015. Green open access

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Abstract

Quantitative measurement of NF-kappa B nuclear translocation is an important research tool in cellular immunology. Established methodologies have a number of limitations, such as poor sensitivity, high cost or dependence on cell lines. Novel imaging methods to measure nuclear translocation of transcriptionally active components of NF-kappa B are being used but are also partly limited by the need for specialist imaging equipment or image analysis software. Herein we present a method for quantitative detection of NF-kappa B rel A nuclear translocation, using immunofluorescence microscopy and the public domain image analysis software ImageJ that can be easily adopted for cellular immunology research without the need for specialist image analysis expertise and at low cost. The method presented here is validated by demonstrating the time course and dose response of NF-kappa B nuclear translocation in primary human macrophages stimulated with LPS, and by comparison with a commercial NF-kappa B activation reporter cell line.

Type: Article
Title: Quantitative imaging assay for NF-kappa B nuclear translocation in primary human macrophages
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.jim.2007.10.015
Publisher version: https://doi.org/10.1016/j.jim.2007.10.015
Language: English
Additional information: © 2007 Elsevier B.V. Open access under CC BY license (https://creativecommons.org/licenses/by/3.0/).
Keywords: Nuclear factor-kappa B, Confocal immunofluoresence, Image analysis, Macrophages
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Infection and Immunity
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > Institute for Global Health
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > Institute for Global Health > Infection and Population Health
URI: https://discovery.ucl.ac.uk/id/eprint/120721

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