Detecting and Characterizing Protein Self-Assembly In Vivo by Flow Cytometry | Zendy

Shriram Venkatesan | Zendy; Tejbir S. Kandola | Zendy; Alejandro Rodríguez Gama | Zendy; Andrew Box | Zendy; Randal Halfmann | Zendy

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Detecting and Characterizing Protein Self-Assembly In Vivo by Flow Cytometry

Author(s) -

Shriram Venkatesan,

Tejbir S. Kandola,

Alejandro Rodríguez Gama,

Andrew Box,

Randal Halfmann

Publication year - 2019

Publication title -

journal of visualized experiments

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.596

H-Index - 91

ISSN - 1940-087X

DOI - 10.3791/59577

Subject(s) - flow cytometry , population , förster resonance energy transfer , in vivo , cytometry , biological system , single cell analysis , microbiology and biotechnology , biophysics , throughput , chemistry , nanotechnology , cell , biology , computational biology , computer science , physics , materials science , biochemistry , fluorescence , genetics , telecommunications , demography , quantum mechanics , sociology , wireless

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.

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