Bifc Assay
Bifc Assay - Related Videos
10:39
Bimolecular Fluorescence Complementation (BiFC) Assay for Protein-Protein Interaction in Onion Cells Using the Helios Gene Gun
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Cited by 18 •
2010
This article illustrates how to properly use the BioRad Helios Gene Gun to introduce plasmid DNA into onion epidermal cells and how to test for protein-protein interactions in onion cells based on the principle of Bimolecular Fluorescence Complementation...
08:21
Detection of Protein Interactions in Plant using a Gateway Compatible Bimolecular Fluorescence Complementation (BiFC) System
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Cited by 30 •
2011
We have developed a technique to test protein-protein interactions in plant. A yellow fluorescent protein (YFP) is split into two non-overlapping fragments. Each fragment is cloned in-frame to a gene of interest via Gateway system, enabling expression of fusion proteins. Reconstitution of YFP signal only occurs when the inquest proteins interact.
14:34
Determination of Tripartite Interaction between Two Monomers of a MADS-box Transcription Factor and a Calcium Sensor Protein by BiFC-FRET-FLIM Assay
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Cited by 3 •
2021
Here we present, a method to visualize ternary complex formation between three protein partners using fluorescent-tagged proteins by BiFC based FRET-FLIM assay. This method is valuable for studying protein-protein interaction complexes in vivo.
12:42
Photoactivated Localization Microscopy with Bimolecular Fluorescence Complementation (BiFC-PALM)
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Cited by 14 •
2015
Protein-protein interactions are visualized in cells with nanometer spatial resolution by combining bimolecular fluorescence complementation (BiFC) with photoactivated localization microscopy (PALM). Described here is the use of BiFC-PALM for imaging Ras-Raf interactions in U2OS cells for visualizing the nanoscale clustering and diffusion of individual Ras-Raf complexes.
08:12
The TUNEL Assay
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2023
One of the hallmarks of apoptosis is the nuclear DNA fragmentation by nucleases. These enzymes are activated by caspases, the family of proteins that execute the cell death program. TUNEL assay is a method that takes advantage of this feature to detect apoptotic cells. In this assay, an enzyme called terminal deoxynucleotidyl transferase catalyzes the addition of dUTP nucleotides to the free 3’ ends of fragmented DNA. By using dUTPs that are labeled with chemical tags that can produce...
FAQs
The two fluorescent protein fragments remain associated with separate fusion partners until the proteins of interest interact. That interaction brings the fragments into close proximity, allowing them to reconstitute a fluorescent signal. The resulting fluorescence provides a visible indication that the paired proteins have come together within the cellular environment being examined.
A BiFC signal does more than indicate that two proteins interact. Its position within the cell can show where that interaction occurs, helping researchers connect molecular binding with cellular organization. This spatial information is particularly useful when examining signaling pathways, protein complexes, or regulatory mechanisms that depend on interactions at specific cellular locations.
By linking fluorescence to the interaction of selected proteins, BiFC can help identify associations that contribute to signaling, complex formation, or regulation. Comparing where the signal appears in cells can provide context for how molecular organization supports a biological process. The method therefore connects protein-level interactions with broader cellular mechanisms.
A typical setup begins by selecting two proteins whose interaction is being investigated and fusing each one to a complementary fluorescent protein fragment. The resulting constructs are examined in cells, where the proteins can encounter one another. Researchers then use microscopy to observe whether fluorescence is produced and where that signal is located.
Microscopy reveals both the presence of a reconstituted fluorescent signal and its distribution within cells. This allows researchers to assess the cellular location of the protein interaction rather than recording only an interaction-dependent outcome. The images can therefore help relate molecular associations to cellular structures, organization, and signaling events.
BiFC is useful when a study needs to visualize protein interactions in their cellular setting. Applications include investigating signaling pathways, characterizing protein complexes, and examining regulatory mechanisms. Its relevance also extends to disease-related research, where altered molecular organization or protein associations may contribute to cellular changes.