DFHBI-1T (solution) 

DFHBI-1T (solution) 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].
Solvent and concentration: DMSO: 20 mM

Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
1. Preparation of stock solution^[1]
It is recommended that the DFHBI-1T stock solution be stored at -20°C or -80°C in the dark after aliquoting.
2. Preparation of working solution
Dilute the stock solution with serum-free medium. The corresponding stock solution can be diluted according to the actual situation. Note that if the solvent is DMSO, the cytotoxicity of DMSO must be considered, and a solvent control should be prepared; if the solvent is pure water, the working solution needs to be filtered and sterilized before adding cells.
Note: Please adjust the concentration of DFHBI-1T working solution according to actual conditions and prepare it before use.
3. Specific staining steps
3.1 Inoculate cells expressing Spinach2-tagged RNA in a suitable culture dish or culture plate and culture under standard cell culture conditions.
3.2 Aspirate the old medium and gently wash the cells twice with pre-warmed PBS to remove residual serum and cell debris.
3.3 Add 20 μM DFHBI-1T working solution to the cells, ensuring complete coverage of the cell monolayer.
3.4 Incubate the cells with the dye for 10 min in a 37°C, 5% CO₂ humidified incubator.
3.5 After incubation, aspirate the dye-containing medium and wash the cells three times with pre-warmed PBS to remove unbound DFHBI-1T.
3.6 Replace PBS with fresh cell culture medium or imaging buffer.
3.7 Immediately image the cells using a fluorescence microscope equipped with a GFP filter set to detect fluorescence of the Spinach2-DFHBI-1T complex.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

320.21

C₁₃H₉F₅N₂O₂

1539318-36-9

Liquid

Light yellow to yellow

O=C1N(C(C)=N/C1=C\C2=CC(F)=C(C(F)=C2)O)CC(F)(F)F

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [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.  [Content Brief]

  [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.  [Content Brief]