| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
DFHBI's primary mechanism of action involves its interaction with specific RNA aptamers, such as Spinach, Spinach2, or Broccoli. The fluorescence of DFHBI is activated upon binding to an RNA aptamer, which restricts the molecule's rotational freedom and suppresses non-radiative decay pathways, such as cis-trans isomerization. This system enables real-time visualization of RNA dynamics, localization, and expression in live cells without requiring protein-based reporters.
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| ln Vitro |
When excited at 447 nm, these RNAs interact with DFHBI to create blue-green fluorescence emission (501 nm). Two RNA aptamers called spinach and spinach2 can be used to genetically encode fluorescent RNA. The fluorescence of DFHBI is bound and activated by Spinach2, which permits imaging of the dynamic location of Spinach2-tagged RNA in living cells. Due to its blue-green fluorescence and lack of optimization for filters typically used in fluorescence microscopy, DFHBI limits the spectral properties of Spinach2. When Spinach and Spinach2 are coupled with DFHBI, their maximum fluorescence excitation is 447 nm, and their peak fluorescence emission is 501 nm [1]. The broccoli-binding fluorophore DFHBI can be used to dye the gel after gel electrophoresis to identify broccoli-labeled RNA in total cellular RNA with selectivity. Spinach is an RNA aptamer, 98 nt in length, that binds to DFHBI and activates its fluorescence. When unbound, DFHBI and spinach are both virtually nonfluorescent, however the complex of DFHBI and spinach fluoresces brightly in both vitro and living cells. DFHBI needs to be shielded from light. It is recommended to store all DFHBI stock solutions in opaque tubes or covered with foil. Wrapped in foil and kept in the dark, plates holding cultures cultured with DFHBI should be kept [2].
In vitro, DFHBI is essentially nonfluorescent when unbound, whereas the Spinach-DFHBI complex is brightly fluorescent. When bound to Spinach2, DFHBI exhibits a peak excitation maximum of 447 nm and a peak fluorescence emission of 501 nm. It displays excitation/emission maxima of 447/501 nm, respectively. |
| ln Vivo |
In vivo, DFHBI is used for imaging RNA in living cells. The Spinach-DFHBI complex is brightly fluorescent in living cells, allowing for the dynamic localization imaging of Spinach2-tagged RNA. DFHBI is cell-permeable, making it suitable for live-cell applications.
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| Enzyme Assay |
In vitro assays for DFHBI typically involve measuring its fluorescence upon binding to RNA aptamers. The probe is incubated with the aptamer, and fluorescence intensity is measured using a fluorometer or plate reader. Excitation and emission spectra are recorded to confirm binding and assess fluorescence activation.
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| Cell Assay |
Cell-based assays for DFHBI involve expressing Spinach2-tagged RNA in living cells and adding the DFHBI probe. Fluorescence is then visualized using fluorescence microscopy or flow cytometry. The dynamic location of the tagged RNA can be monitored in real-time.
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| Animal Protocol |
In vivo animal studies for DFHBI are limited, as the compound is primarily used in cell culture systems. Its use in live animals for RNA imaging has not been extensively explored.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of DFHBI are not relevant for therapeutic applications, as it is a fluorescent probe. The compound has a molecular weight of 252.22 g/mol. It is typically dissolved in DMSO for in vitro experiments.
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| Toxicity/Toxicokinetics |
The toxicological profile of DFHBI has not been extensively characterized. As a research compound, it should be handled with appropriate safety precautions. Its use is limited to research applications.
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| References | |
| Additional Infomation |
DFHBI is a mimic of green fluorescent protein (GFP) fluorophore for imaging RNA in living cells. Fluorescence is activated by binding to Spinach2 or Broccoli aptamers. It is a valuable tool for studying RNA dynamics and localization. It is not approved for therapeutic use. While DFHBI serves as the foundational fluorophore for Spinach aptamer-based RNA imaging, its derivatives DFHBI-1T and BI demonstrate quantifiably distinct performance profiles that directly impact experimental outcomes.
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| Molecular Formula |
C12H10F2N2O2
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|---|---|
| Molecular Weight |
252.216809749603
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| Exact Mass |
252.071
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| CAS # |
1241390-29-3
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| PubChem CID |
70808995
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
326.2±52.0 °C at 760 mmHg
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| Flash Point |
151.1±30.7 °C
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| Vapour Pressure |
0.0±0.7 mmHg at 25°C
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| Index of Refraction |
1.579
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| LogP |
1.66
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
18
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| Complexity |
410
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=N/C(=C\C2=CC(=C(C(=C2)F)O)F)/C(=O)N1C
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| InChi Key |
ZDDIJYXDUBFLID-YHYXMXQVSA-N
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| InChi Code |
InChI=1S/C12H10F2N2O2/c1-6-15-10(12(18)16(6)2)5-7-3-8(13)11(17)9(14)4-7/h3-5,17H,1-2H3/b10-5-
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| Chemical Name |
(5Z)-5-[(3,5-difluoro-4-hydroxyphenyl)methylidene]-2,3-dimethylimidazol-4-one
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ≥ 83.33 mg/mL (~330.39 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.43 mg/mL (5.67 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 14.3 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1.43 mg/mL (5.67 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 14.3 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 1.43 mg/mL (5.67 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.9648 mL | 19.8240 mL | 39.6479 mL | |
| 5 mM | 0.7930 mL | 3.9648 mL | 7.9296 mL | |
| 10 mM | 0.3965 mL | 1.9824 mL | 3.9648 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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