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Rapid flow cytometric measurement of protein inclusions and nuclear trafficking.

Sci Rep.. 2016-08; 
Whiten DR, San Gil R, McAlary L, Yerbury JJ, Ecroyd H, Wilson MR.
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PCR Cloning and Subcloning ... SOD1-tdTomato constructs were created by replacing the GFP sequences in the SOD1-GFP plasmids with tdTomato (Genscript) 18 . The expression vector pCMV6-AC-GFP containing FUS was obtained from Origene and site ... Get A Quote

摘要

Proteinaceous cytoplasmic inclusions are an indicator of dysfunction in normal cellular proteostasis and a hallmark of many neurodegenerative diseases. We describe a simple and rapid new flow cytometry-based method to enumerate, characterise and, if desired, physically recover protein inclusions from cells. This technique can analyse and resolve a broad variety of inclusions differing in both size and protein composition, making it applicable to essentially any model of intracellular protein aggregation. The method also allows rapid quantification of the nuclear trafficking of fluorescently labelled molecules.

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