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Protein Dynamics in Living Cells

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...

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Related Experiment Video

Updated: Jun 4, 2026

Protein-protein Interactions Visualized by Bimolecular Fluorescence Complementation in Tobacco Protoplasts and Leaves

Protein-protein Interactions Visualized by Bimolecular Fluorescence Complementation in Tobacco Protoplasts and Leaves

Published on: March 9, 2014

In Planta Visualization of Protein Interactions Using Bimolecular Fluorescence Complementation (BiFC).

Rainer Waadt¹, Jörg Kudla

¹Institut für Botanik und Botanischer Garten, Universität Münster, 48149 Münster, Germany.

|March 2, 2011

Summary

Bimolecular fluorescence complementation (BiFC) visualizes protein interactions in living cells. This method uses Agrobacterium-mediated transient expression in Nicotiana benthamiana for high-efficiency protein complex analysis.

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Published on: September 16, 2011

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Last Updated: Jun 4, 2026

Protein-protein Interactions Visualized by Bimolecular Fluorescence Complementation in Tobacco Protoplasts and Leaves

Protein-protein Interactions Visualized by Bimolecular Fluorescence Complementation in Tobacco Protoplasts and Leaves

Published on: March 9, 2014

Bimolecular Fluorescence Complementation

Bimolecular Fluorescence Complementation

Published on: April 15, 2011

Detection of Protein Interactions in Plant using a Gateway Compatible Bimolecular Fluorescence Complementation (BiFC) System

Detection of Protein Interactions in Plant using a Gateway Compatible Bimolecular Fluorescence Complementation (BiFC) System

Published on: September 16, 2011

Area of Science:

Molecular Biology
Cell Biology
Biochemistry

Background:

Bimolecular fluorescence complementation (BiFC) is a powerful technique for visualizing protein-protein interactions in live cells.
It relies on the reassembly of fluorescent protein fragments (e.g., enhanced yellow fluorescent protein) when brought together by interacting proteins.

Purpose of the Study:

To describe a robust protocol for BiFC assays using Agrobacterium-mediated transient expression in Nicotiana benthamiana leaf cells.
To enable visualization and colocalization analysis of protein complexes in plant cells.

Main Methods:

Agrobacterium-mediated transient expression in Nicotiana benthamiana leaf cells.
Fusion of interacting proteins to fragments of enhanced yellow fluorescent protein (eYFP).
Analysis of fluorescence complementation to confirm protein interactions and localization.

Main Results:

Achieved high transformation rates (up to 90%) in Nicotiana benthamiana leaf cells.
Enabled simultaneous expression and colocalization analysis of multiple proteins.
Facilitated investigation of protein interactions at various time points and in different cell types.

Conclusions:

The described Agrobacterium-mediated transient expression system is highly efficient for BiFC assays in Nicotiana benthamiana.
This method allows detailed cellular localization studies of protein complexes.
It offers advantages for studying toxic proteins and comparative interaction analyses.