3, 5-difluoro-4-hydroxybenzylidene imidazolinone ( DFHBI )

- Overview
  - 3'5'-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI) is a non-fluorescent small molecule that resembles the chromophore of green fluorescent protein (GFP). Upon binding to the "Spinach™" aptamer, DFHBI is converted to a highly fluorescent state that can be detected at the emission wavelength of 501 nm. DFHBI is cell permeable with negligible toxicity in living cells. DFHBI can be used to label any genetically encoded Spinach™ family of RNA tags that include Spinach™, Spinach2™, and Broccoli™. Live-cell imaging of tagged RNA can be performed using a standard fluorescence microscopy. Further, DFHBI can also selectively detect tagged RNA in total RNA gel electrophoresis. Thus, bypassing the need for Northern blot analysis.
    
    Data
    
    Figure 1. HEK293T cells were transfected with either 5S-control aptamer or 5S-Spinach™ plasmids and incubated with 20 µM DFHBI. Images are phase overlay with Hoechst-stained nuclei (blue) and Spinach™ fluorescence (green).
    
    Figure 2. Spinach™ RNA robustly activates the fluorescence of DFHBI. Each tube contained the indicated solution and was irradiated at 465 nm.
    
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    More Details
    
    Fluorophores for Imaging RNA in Living Cells
- Properties
  - Categories
    
    Fluorophores
    
    Storage
    
    Stable in the dark for 2 year at -20 °C.
    
    Alternative Name
    
    DFHBI
    
    CAS Number
    
    1241390-29-3
    
    Molecular Formula
    
    C12H10F2N2O2
    
    Molecular Weight
    
    252.22
    
    Appearance
    
    Lyophilized dye
    
    Other Properties
    
    Products are analyzed by 1H NMR and LC-MS and provided at purity of >95% by HPLC.
    Absorption and fluorescence emission spectra of DFHBI in pH 7.4 buffer.
    
    * This product is intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
- Applications
  - Application Description
    
    DFHBI can be used to label any genetically encoded spinach RNA tag family, including Spinach, Spinach2 and Broccoli. Live-cell imaging of tagged RNA can be performed using standard fluorescence microscopy.
- Reference
  - RNA mimics of green fluorescent protein. Paige JS, Wu KY, Jaffrey SR. Science. 2011 Jul 29;333(6042):642-6.
    Fluorescence imaging of cellular metabolites with RNA. Paige JS, Nguyen-Duc T, Song W, Jaffrey SR. Science. 2012 Mar 9;335(6073):1194.