Name: "3, 5-difluoro-4-hydroxybenzylidene imidazolinone ( DFHBI )"
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"3, 5-difluoro-4-hydroxybenzylidene imidazolinone ( DFHBI )" | Amerigo Scientific

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./

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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)./

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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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