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Single-Molecule Imaging of Nuclear Transport
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Protocol for single-molecule fluorescence recovery after photobleaching microscopy to analyze the dynamics and

Mark Tingey1, Yichen Li1, Weidong Yang1

  • 1Department of Biology Temple University, Philadelphia, PA 19122, USA.

STAR Protocols
|May 19, 2021
PubMed

Keywords:
Atomic Force Microscopy (AFM)Biotechnology and bioengineeringMicrobiologyMolecular Biology

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Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...