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DFHBI

Cat No.:V19702 Purity: ≥98%
DFHBI is a mimic of green fluorescent protein (GFP) fluorophore for imaging RNA in living cells.
DFHBI
DFHBI Chemical Structure CAS No.: 1241390-29-3
Product category: New1
This product is for research use only, not for human use. We do not sell to patients.
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5mg
10mg
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Purity & Quality Control Documentation

Purity: ≥98%

Product Description

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. Exhibits peak excitation maxima of 447 nm and peak fluorescence emission of 501 nm when bound to Spinach2.

Biological Activity I Assay Protocols (From Reference)
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].
References

[1]. Plug-and-play fluorophores extend the spectral properties of Spinach. J Am Chem Soc. 2014 Jan 29;136(4):1198-201.

[2]. Using Spinach-based sensors for fluorescence imaging of intracellular metabolites and proteins in living bacteria. Nat Protoc. 2014 Jan;9(1):146-55.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C12H10F2N2O2
Molecular Weight
252.216809749603
Exact Mass
252.071
CAS #
1241390-29-3
PubChem CID
70808995
Appearance
Light yellow to yellow solid powder
Density
1.4±0.1 g/cm3
Boiling Point
326.2±52.0 °C at 760 mmHg
Flash Point
151.1±30.7 °C
Vapour Pressure
0.0±0.7 mmHg at 25°C
Index of Refraction
1.579
LogP
1.66
Hydrogen Bond Donor Count
1
Hydrogen Bond Acceptor Count
5
Rotatable Bond Count
1
Heavy Atom Count
18
Complexity
410
Defined Atom Stereocenter Count
0
SMILES
CC1=N/C(=C\C2=CC(=C(C(=C2)F)O)F)/C(=O)N1C
InChi Key
ZDDIJYXDUBFLID-YHYXMXQVSA-N
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-
Chemical Name
(5Z)-5-[(3,5-difluoro-4-hydroxyphenyl)methylidene]-2,3-dimethylimidazol-4-one
HS Tariff Code
2934.99.9001
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)
Solubility Data
Solubility (In Vitro)
DMSO : ≥ 83.33 mg/mL (~330.39 mM)
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.

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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.
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 corn oil and mix evenly.


 (Please use freshly prepared in vivo formulations for optimal results.)
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.

Calculator

Molarity Calculator allows you to calculate the mass, volume, and/or concentration required for a solution, as detailed below:

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An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
  • Enter 350.26 in the Molecular Weight (MW) box
  • Enter 10 in the Concentration box and choose the correct unit (mM)
  • Enter 5 in the Volume box and choose the correct unit (mL)
  • Click the “Calculate” button
  • The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
  • Enter 10 into the Concentration (Start) box and choose the correct unit (mM)
  • Enter 25 into the Concentration (End) box and select the correct unit (mM)
  • Enter 25 into the Volume (End) box and choose the correct unit (mL)
  • Click the “Calculate” button
  • The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box
g/mol

Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
Instructions to calculate molar mass (molecular weight) of a chemical compound:
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Definitions of molecular mass, molecular weight, molar mass and molar weight:
  • Molecular mass (or molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
  • Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.
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  • The answer appears in the Volume (to add to vial) box
In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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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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