DFHBI is a small molecule that resembles the chromophore of green fluorescent protein (GFP). Spinach and DFHBI are essentially nonfluorescent when unbound, whereas the Spinach-DFHBI complex is brightly fluorescent both in vitro and in living cells.
体外研究 (In Vitro)
These RNAs interact with DFHBI to produce a bluish-green fluorescence emission (501 nm) after excitation at 447 nm. Spinach and Spinach2 are RNA aptamers that can be used for the genetic encoding of fluorescent RNA. Spinach2 binds and activates the fluorescence of DFHBI, allowing the dynamic localizations of Spinach2-tagged RNAs to be imaged in live cells. The spectral properties of Spinach2 are limited by DFHBI, which produces fluorescence that is bluish-green and is not optimized for filters commonly used in fluorescence microscopes. Spinach and Spinach2 bind to DFHBI have fluorescence excitation maxima of 447 nm and peak fluorescence emission of 501 nm[1]. Broccoli-tagged RNAs are selectively detected in total cellular RNA by gel electrophoresis followed by staining of gels with DFHBI, the Broccoli-binding fluorophore. Spinach is a 98-nt-long RNA aptamer that binds to and switches on the fluorescence of DFHBI. Both Spinach and DFHBI are essentially nonfluorescent when unbound, whereas the Spinach-DFHBI complex is brightly fluorescent both in vitro and in living cells. DFHBI should be shielded from light. All stock solutions of DFHBI should be maintained in dark tubes or wrapped in foil. Plates containing cultures incubated with DFHBI should be kept in the dark by using a foil overwrap[2].
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量
252.22
Formula
C12H10F2N2O2
CAS 号
1241390-29-3
运输条件
Room temperature in continental US; may vary elsewhere.
[1]. Song W, et al. Plug-and-play fluorophores extend the spectral properties of Spinach. J Am Chem Soc. 2014 Jan 29;136(4):1198-201.
[2]. Strack RL, et al. Using Spinach-based sensors for fluorescence imaging of intracellular metabolites and proteins in living bacteria. Nat Protoc. 2014 Jan;9(1):146-55.
DFHBI-1T is a membrane-permeable RNA aptamers-activated fluorescence probe (ex/em=472 nm/507 nm). DFHBI-1T binds to RNA aptamers (Spinach, Spinach2, iSpinach, and Broccoli) and causes specific fluorescence and lower background fluorescence. DFHBI-1T is used to image RNA in live cells[1][2].
体外研究 (In Vitro)
DFHBI-1T (20 μM; for 10 min) increases fluorescence in COS7 cells expressing (CGG)60-Spinach2 over DFHBI (20 μM)[1]. Broccoli-DFHBI-1T has ex/em=472 nm/507 nm and Spinach2-DFHBI-1T has ex/em=482 nm/505 nm[2].
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量
320.21
Formula
C13H9F5N2O2
CAS 号
1539318-36-9
运输条件
Room temperature in continental US; may vary elsewhere.
[1]. Wenjiao Song, et al. Plug-and-play fluorophores extend the spectral properties of Spinach. J Am Chem Soc. 2014 Jan 29;136(4):1198-201.
[2]. Grigory S Filonov, et al. Broccoli: rapid selection of an RNA mimic of green fluorescent protein by fluorescence-based selection and directed evolution. J Am Chem Soc. 2014 Nov 19;136(46):16299-308.