4-(3,5-difluoro-4-hydroxybenzylidene)-2-methyl-1-(2,2,2-trifluoroethyl)-1H-imidazol-5(4 H)-one ( DFHBI-1T )

- Overview
  - DFHBI-1T is a non-fluorescent dye that consists of an 1,1,1-trifluoroethyl substituent on the imidazolone ring of the DFHBI fluorophore. Upon binding to the Spinach™ aptamer family, DFHBI-1T is converted to a highly fluorescent state that can be detected at the emission wavelength of 505 nm. DFHBI-1T is cellpermeable with negligible toxicity in living cells and can be used to label any genetically encoded Spinach™, Spinch2™, and Broccoli™ RNA tags. DFHBI-1T has been found to have lower background fluorescence than DFHBI and exhibits an overall increase in brightness in living cells. Further, DFHBI-1T can also selectively detect tagged RNA in total RNA gel electrophoresis. Thus, bypassing the need for Northern blot analysis.
    
    Data
    
    A. Live-cell imaging of a COS7 cell expressing CGG60-Spinach2™ in the presence of either 20 µM DFHBI or DFHBI-1T. Images of the same cell were acquired using a 100 msec exposure with a GFP filter set. B. Quantification of average DFHBI-1T brightness of 10 cells expressing CGG60-Spinach™ normalized to the brightness of DFHBI.
    
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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-1T
    
    CAS Number
    
    1539318-36-0
    
    Molecular Formula
    
    C13H9F5N2O2
    
    Molecular Weight
    
    320.21
    
    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 Spinach2™/DFHBI-1T in pH 7.4 buffer.
    Excitation maximum: 482 nm
    Emission maximum: 505 nm
    
    * 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-1T can be bound to Spinach aptamers or Spinach-derived aptamers, such as Spinch2 and Broccoli, for imaging RNA in living cells
- 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.