DFHBI-1T

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]. In Vitro:Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
1.1 Preparation of stock solution^[1]
Use anhydrous DMSO to prepare a 50 mM DFHBI-1T stock solution.
Note: It is recommended that the DFHBI-1T stock solution be stored at -20℃ or -80℃ in the dark after aliquoting.
1.2 Preparation of working solution
Dilute the stock solution with serum-free medium to prepare a 20 μM DFHBI-1T working solution.
Note: Please adjust the concentration of DFHBI-1T working solution according to actual conditions and prepare it before use.
2 Specific staining steps
1) Inoculate cells expressing Spinach2-tagged RNA in a suitable culture dish or culture plate and culture under standard cell culture conditions. 2) Aspirate the old medium and gently wash the cells twice with pre-warmed PBS to remove residual serum and cell debris.
3) Add 20 μM DFHBI-1T working solution to the cells, ensuring complete coverage of the cell monolayer.
4) Incubate the cells with the dye for 10 min in a 37°C, 5% CO₂ humidified incubator.
5) After incubation, aspirate the dye-containing medium and wash the cells three times with pre-warmed PBS to remove unbound DFHBI-1T.
6) Replace PBS with fresh cell culture medium or imaging buffer.
7) Immediately image the cells using a fluorescence microscope equipped with a GFP filter set to detect fluorescence of the Spinach2-DFHBI-1T complex.