| Chapter title |
Bimolecular Fluorescence Complementation (BiFC) Analysis of Protein-Protein Interactions and Assessment of Subcellular Localization in Live Cells.
|
|---|---|
| Chapter number | 9 |
| Book title |
High-Resolution Imaging of Cellular Proteins
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-6352-2_9 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6350-8, 978-1-4939-6352-2
|
| Authors |
Evan P. S. Pratt, Jake L. Owens, Gregory H. Hockerman, Chang-Deng Hu |
| Editors |
Steven D. Schwartzbach, Omar Skalli, Thomas Schikorski |
| Abstract |
Bimolecular fluorescence complementation (BiFC) is a fluorescence imaging technique used to visualize protein-protein interactions (PPIs) in live cells and animals. One unique application of BiFC is to reveal subcellular localization of PPIs. The superior signal-to-noise ratio of BiFC in comparison with fluorescence resonance energy transfer or bioluminescence resonance energy transfer enables its wide applications. Here, we describe how confocal microscopy can be used to detect and quantify PPIs and their subcellular localization. We use basic leucine zipper transcription factor proteins as an example to provide a step-by-step BiFC protocol using a Nikon A1 confocal microscope and NIS-Elements imaging software. The protocol given below can be readily adapted for use with other confocal microscopes or imaging software. |
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