Bimolecular Fluorescence Complementation (BiFC) Service

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Bimolecular Fluorescence Complementation (BiFC) Service

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CD BioSciences provides clients with bimolecular fluorescence complementation (BiFC) systems to help verify the existence of protein interactions and detect interaction localization. Our service can also be applied to drug development by the construction of BiFC-stable cell lines to screen drugs with potential therapeutic functions.

BiFC technology is capable of determining the localization and interaction of proteins in cells. The principle is that when interacting proteins are attached to and co-expressed with two non-fluorescently fragments, the fragments are brought close to each other, and active fluorescent groups are re-formed. By observing the fluorescence through the microscope, it can determine whether the bait protein interacts with the prey protein.

Formation of active pattern recognition receptor complexes. Figure 1. The principle of BiFC. (Miller, K. E., et al., 2015)

Service Content

Experimental content

  • Construction of plasmids containing target proteins. (clients need to confirm whether the target protein is attached to the N- or C-terminus of the fluorescent protein)

    • a) Gene optimization.

    b) Subcloning design.

    c) Synthesize gene.

    d) Subcloning into vector.

  • Recombinant target plasmid transformation of Agrobacterium.
  • Injection of Agrobacterium sap into tobacco leaves.
  • Confocal microscope observation and photography.

Fluorescent proteins we provide for the BiFC system

GFP (green fluorescent protein) BFP (blue fluorescent protein) Citrine
YFP (yellow fluorescent protein) RFP (red fluorescent protein) Cerulean
CFP (cyan fluorescent protein) Venus mCherry

BiFC extension systems

  • Multi-color fluorescence complementary technology

We offer multicolor fluorescent complementation technology that enables the simultaneous detection of multiple sets of protein interactions in a cell and also allows comparison of the intensity of protein interactions by the strength of the fluorescence.

  • BiFC-FRET technology

We combine BiFC and FRET technology to establish BiFC-FRET, which can detect the interactions between three proteins.

  • BiFC-YTH technology

We combine the BiFC system based on GFP and YFP with yeast two-hybrid technology to study the self-interaction of viruses in plant cells.

Advantages and disadvantages of BiFC technology

Advantages Disadvantages
  • Visualization.
  • Can be applied to different hosts.
  • Can be applied in vivo and in vitro.
  • Clean background, more sensitive detection.
  • Low instrumentation requirements and cost control.
  • Sensitive to temperature.
  • Information lag, unable to observe interactions in real-time.

Materials to be Provided by Clients

Sample type Requirement Details
Plasmids Volume > 20µl.
> 20ng/µl, free of impurities.
OD (260/280) is 1.6-2.0. 		Sequencing results.
Empty control plasmid.
Escherichia coli glycerol > 500µl. Sequencing results.
Empty control bacterial solution.
Bacterial solution resistance.
Bacteria with better activity within three months.
Seeds Dicotyledon > 50 seeds.
Monocotyledon > 25 seeds. 		Seeds within three months.
Germination rate >90%.

Delivery Reports

  • Full sequence information and annotation information of recombinant vectors.
  • Enzymatic mapping and sequencing results of the recombinant vector.
  • Recombinant vector, including one copy of plasmid and one copy of E. coli bacterial fluid (25% glycerol bacteria).
  • Original pictures (fluorescence channel maps, chloroplast autofluorescence channel maps, bright field, superimposed maps, rice protoplast without chloroplast autofluorescence channel maps).
  • The complete experimental report, including detailed experimental steps and instrumental parameters.

CD BioSciences is a leading biological company for plant protein research. Our team of experienced scientists uses cutting-edge fluorescence microscopy and image analysis techniques to visualize and characterize protein interactions in living plant cells. Whether you are studying signaling pathways, protein localization, or molecular interactions in plants, contact us to help you capture and analyze the complex dynamics of protein complexes.

Reference

  1. Miller, K. E., et al. (2015). Bimolecular Fluorescence Complementation (BiFC) Analysis: Advances and Recent Applications for Genome-Wide Interaction Studies. Journal of molecular biology. 427(11), 2039-2055.

For research use only, not for clinical use.