Detecting and Characterizing Protein Self-Assembly In Vivo by Flow Cytometry
| Autor: | Andrew C. Box, Shriram Venkatesan, Randal Halfmann, Tejbir S. Kandola, Alejandro Rodríguez-Gama |
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| Rok vydání: | 2019 |
| Předmět: |
0303 health sciences
Protein function medicine.diagnostic_test General Immunology and Microbiology General Chemical Engineering General Neuroscience Proteins In Vitro Techniques Protein aggregation Flow Cytometry Protein subcellular localization prediction Article General Biochemistry Genetics and Molecular Biology Flow cytometry 03 medical and health sciences 0302 clinical medicine Förster resonance energy transfer Proteostasis In vivo medicine Biophysics Humans Self-assembly 030217 neurology & neurosurgery 030304 developmental biology |
| Zdroj: | J Vis Exp |
| ISSN: | 1940-087X |
| DOI: | 10.3791/59577-v |
| Popis: | Protein self-assembly governs protein function and compartmentalizes cellular processes in space and time. Current methods to study it suffer from low-sensitivity, indirect read-outs, limited throughput, and/or population-level rather than single-cell resolution. We designed a flow cytometry-based single methodology that addresses all of these limitations: Distributed Amphifluoric FRET or DAmFRET. DAmFRET detects and quantifies protein self-assemblies by sensitized emission FRET in vivo, enables deployment across model systems-from yeast to human cells-and achieves sensitive, single-cell, high-throughput read-outs irrespective of protein localization or solubility. |
| Databáze: | OpenAIRE |
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