Abstract
Immunofluorescence can be a powerful tool to detect protein levels, intracellular localization, and post-translational modifications. However, standard immunofluorescence provides only a still picture and thus lacks temporal information. Here, we describe a method to extract temporal information from immunofluorescence images of fixed cells. In addition, we provide an optional protocol that uses micropatterns, which increases the accuracy of the method. These methods allow assessing how protein levels, intracellular localization, and post-translational modifications change through the cell cycle.
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Acknowledgements
This work was supported by the Swedish Research Council, the Swedish Foundation for Strategic Research, and the Swedish Cancer Society. We thank all members of the Lindqvist Lab for discussions.
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Akopyan, K., Lindqvist, A., Müllers, E. (2016). Cell Cycle Dynamics of Proteins and Post-translational Modifications Using Quantitative Immunofluorescence. In: Coutts, A., Weston, L. (eds) Cell Cycle Oscillators. Methods in Molecular Biology, vol 1342. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2957-3_9
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DOI: https://doi.org/10.1007/978-1-4939-2957-3_9
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2956-6
Online ISBN: 978-1-4939-2957-3
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