| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg |
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| 10mg | |||
| Other Sizes |
| Targets |
Spinach2 RNA aptamer [1][2]
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|---|---|
| ln Vitro |
When bound to Spinach2, DFHBI-2T exhibited a marked 53 nm red shift in the excitation peak (from 447 nm to 500 nm) and a 22 nm red shift in the emission peak (from 501 nm to 523 nm) compared to DFHBI. The overall brightness was reduced due to a decrease in quantum yield. [1]
The photophysical properties of the Spinach2-DFHBI-2T complex were: maximum absorption at 460 nm, maximum excitation at 500 nm, maximum emission at 523 nm, extinction coefficient of 29,000 M⁻¹ cm⁻¹, quantum yield of 0.12, brightness of 22 (relative to Spinach2-DFHBI), and a dissociation constant (Kd) of approximately 1,300 nM (1.2 μM). [1] The free DFHBI-2T fluorophore (not bound to Spinach2) exhibited: maximum absorption at 460 nm, maximum emission at 515 nm, extinction coefficient of 34,800 M⁻¹ cm⁻¹, and a fluorescence quantum yield of 0.0012. [1] In vitro, incubation of the Spinach2-based S-adenosylmethionine (SAM) sensor (composed of Spinach2 and the SAM aptamer) with DFHBI-2T resulted in SAM-dependent fluorescence emission at 523 nm. [1] Fluorescence observable through the yellow emission channel is produced by spinach2 imaged DFHBI-2T[2]. |
| ln Vivo |
In COS7 cells expressing (CGG)₆₀-Spinach2, when the media was switched to media containing 20 μM DFHBI-2T, fluorescence was markedly reduced when imaging with the GFP filter cube but was readily detectable using the YFP filter cube. This indicates that Spinach2 imaged with DFHBI-2T results in fluorescence detectable using the yellow emission channel. [1]
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| Enzyme Assay |
Spectra collection for Spinach2-DFHBI-2T complex: Excitation and emission spectra were collected in the presence of excess Spinach2 (5 μM) with 1 μM DFHBI-2T in binding buffer (40 mM HEPES, pH 7.4, 125 mM KCl, 10 mM MgCl₂). Excitation spectra were collected at the emission maximum wavelength, and emission spectra were collected at the excitation maximum wavelength. [1]
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| Cell Assay |
Live-cell imaging of Spinach2 fusion RNAs with DFHBI-2T: COS7 cells expressing (CGG)₆₀-Spinach2 were cultured in media containing either DFHBI or DFHBI-2T. Cells were imaged using both GFP and YFP filter sets. Scale bar, 10 μm. [1]
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| References | |
| Additional Infomation |
DFHBI-2T ((Z)-4-(3,5-difluoro-4-hydroxybenzylidene)-2-methyl-1-(2,2,2-trifluoroethyl)-1H-imidazol-5(4H)-one) is a fluorophore that binds to and is activated by the Spinach2 RNA aptamer. It was synthesized by switching the methyl substituent at the C-2 position of the imidazolinone ring in DFHBI to a trifluoromethyl group. [1]
When bound to Spinach2, DFHBI-2T exhibits spectral properties that are more compatible with YFP filter cubes (excitation bandpass filter transmitting 500 ± 10 nm light, dichroic mirror at 515 nm, and emission filter transmitting 535 ± 15 nm light) rather than standard GFP filter cubes. This allows Spinach2-tagged constructs to be imaged using the YFP filter cube. [1] The bulky trifluoromethyl moiety at the C-2 position may exhibit steric hindrance with the Spinach2 aptamer, as evidenced by the increase in Kd to approximately 1.2 μM. [1] DFHBI-2T is part of a "plug-and-play" system for RNA imaging in living cells, allowing the spectral properties of Spinach2 to be altered based on specific experimental needs. This can be valuable for experiments in which simultaneous imaging of Spinach2-tagged RNAs and CFP-tagged proteins is desired. [1] |
| Molecular Formula |
C12H7F5N2O2
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|---|---|
| Molecular Weight |
306.188200235367
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| Exact Mass |
306.04
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| CAS # |
1539318-40-5
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| Related CAS # |
DFHBI-1T;1539318-36-9
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| PubChem CID |
129080921
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| Appearance |
Light yellow to yellow solid powder
|
| LogP |
2.5
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
21
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| Complexity |
490
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1(C(F)(F)F)=N/C(=C\C2=CC(F)=C(O)C(F)=C2)/C(=O)N1C
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| InChi Key |
CGFZNPPTJGGWPN-YWEYNIOJSA-N
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| InChi Code |
InChI=1S/C12H7F5N2O2/c1-19-10(21)8(18-11(19)12(15,16)17)4-5-2-6(13)9(20)7(14)3-5/h2-4,20H,1H3/b8-4-
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| Chemical Name |
(5Z)-5-[(3,5-difluoro-4-hydroxyphenyl)methylidene]-3-methyl-2-(trifluoromethyl)imidazol-4-one
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| Synonyms |
DFHBI-2T; DFHBI 2T; 1539318-40-5; (5Z)-5-[(3,5-difluoro-4-hydroxyphenyl)methylidene]-3-methyl-2-(trifluoromethyl)imidazol-4-one; SCHEMBL18797644; CGFZNPPTJGGWPN-YWEYNIOJSA-N;
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.2659 mL | 16.3297 mL | 32.6595 mL | |
| 5 mM | 0.6532 mL | 3.2659 mL | 6.5319 mL | |
| 10 mM | 0.3266 mL | 1.6330 mL | 3.2659 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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