Characterization of aggregate load and pattern in living yeast cells by flow cytometry
Protein aggregation is both a hallmark of and a driving force for a number of diseases. It is therefore important to identify the nature of these aggregates and the mechanism(s) by which the cell counteracts their detrimental properties. Currently, the study of aggregation in vivo is performed prima...
Gespeichert in:
| Hauptverfasser: | , , , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
1 September 2016
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| In: |
BioTechniques
Year: 2016, Jahrgang: 61, Heft: 3, Pages: 137-148 |
| ISSN: | 1940-9818 |
| Online-Zugang: | Verlag, Volltext: https://www.biotechniques.com/BiotechniquesJournal/2016/September/Characterization-of-aggregate-load-and-pattern-in-living-yeast-cells-by-flow----cytometry/biotechniques-365063.html
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| Verfasserangaben: | Itahisa Hernández Hidalgo, Thomas Fleming, Volker Eckstein, Stephan Herzig, Peter P. Nawroth, Jens Tyedmers |
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| 520 | |a Protein aggregation is both a hallmark of and a driving force for a number of diseases. It is therefore important to identify the nature of these aggregates and the mechanism(s) by which the cell counteracts their detrimental properties. Currently, the study of aggregation in vivo is performed primarily using fluorescently tagged versions of proteins and analyzing the aggregates by fluorescence microscopy. While this strategy is considered the gold standard, it has several limitations, particularly with respect to its suitability for high-throughput screening (HTS). Here, using a GFP fusion of the well-characterized yeast prion amyloid protein [PSI+], we demonstrate that flow cytometry, which utilizes the same physical principles as fluorescence microscopy, can be used to determine the aggregate load and pattern in live and fixed yeast cells. Furthermore, our approach can easily be applied to high-throughput analyses such as screenings with a yeast deletion library. | ||
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| 650 | 4 | |a flow cytometry | |
| 650 | 4 | |a Flow Cytometry | |
| 650 | 4 | |a fluorescence activated cell sorting (FACS) | |
| 650 | 4 | |a Green Fluorescent Proteins | |
| 650 | 4 | |a high-throughput screening (HTS) | |
| 650 | 4 | |a High-Throughput Screening Assays | |
| 650 | 4 | |a Peptide Termination Factors | |
| 650 | 4 | |a Protein Aggregates | |
| 650 | 4 | |a protein aggregation | |
| 650 | 4 | |a Recombinant Fusion Proteins | |
| 650 | 4 | |a Saccharomyces cerevisiae | |
| 650 | 4 | |a Saccharomyces cerevisiae Proteins | |
| 650 | 4 | |a yeast prion | |
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