Abstract
Pulse shape analysis (PulSA) is a flow cytometry-based method that can be used to study protein localization patterns in cells. Examples for its use include tracking the formation of inclusion bodies of polyglutamine-expanded proteins and other aggregating proteins. The method can also be used for phenomena relating to protein movements in cells such as translocation from the cytoplasm to the nucleus, trafficking from the plasma membrane to the Golgi, and stress granule formation. An attractive feature is its capacity to quantify these parameters in whole-cell populations very quickly and in high throughput. We describe the basic experimental details for performing PulSA using expression of GFP-tagged proteins, endogenous proteins labelled immunofluorescently, and organelle dyes.
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Acknowledgements
This work was funded by grants to D.M.H. from the Australian Research Council (DP120102763) and the Hereditary Disease Foundation. D.M.H. is a Grimwade Research fellow, funded by the Miegunyah Trust.
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Ramdzan, Y.M., Wood, R., Hatters, D.M. (2013). Pulse Shape Analysis (PulSA) to Track Protein Translocalization in Cells by Flow Cytometry: Applications for Polyglutamine Aggregation. In: Hatters, D., Hannan, A. (eds) Tandem Repeats in Genes, Proteins, and Disease. Methods in Molecular Biology, vol 1017. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-438-8_6
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DOI: https://doi.org/10.1007/978-1-62703-438-8_6
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