Dibenzylfluorescein
Based on 1 Customer Validation
Dibenzylfluorescein (DBF) is a fluorogenic probe (Fluoresecent dye) that acts as a substrate for specific cytochrome P450 (CYP) isoforms, including CYP3A4, CYP2C8, CYP2C9, CYP2C19, and aromatase (CYP19). Dibenzylfluorescein is typically used near its Km value of 0.87-1.9 μM (Ex=485 nm，Em=535 nm). Dibenzylfluorescein is used to detect changes in CYP catalytic activity caused by drugs or disease.
For research use only. We do not sell to patients.
- Purity: 99.54%
- CAS No.: 97744-44-0
- Formula: C34H24O5
- Molecular Weight:512.55
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Storage:
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Purity & Documentation
- References
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Help & FAQs
Biological Activity
Guidelines (The following is our recommended protocol. This protocol is provided for reference only and should be modified according to your specific needs).
Experimental Method for P450-Catalyzed Dibenzylfluorescein Metabolism and the Effect of Alkali[3]:
Reaction process: Dibenzylfluorescein is dealkylated by P450 to generate fluorescein benzyl ester, which can be hydrolyzed into fluorescein in the presence of sodium hydroxide. Addition of 2 M sodium hydroxide also causes Dibenzylfluorescein to decompose into fluorescein benzyl ether.
1. The reaction conditions for CYP2C19 enzyme activity and inhibition assays in a 150 μL reaction system were as follows: 0.1 M Tris-HCl buffer (pH 7.4), 10 μM Dibenzylfluorescein, 15 pmol CYP2C19 enzyme, and 50 μL NADPH regenerating system. The NADPH regenerating system contained 1.13 mM NADP, 12.5 mM isocitrate, 56.33 mM KCl, 187.5 mM Tris-HCl, pH 7.4, 12.5 mM MgCl2, 0.0125 mM MnCl2, and 0.075 U/mL isocitrate dehydrogenase.
2. Samples were incubated at 37°C for 30-60 minutes. The reaction was terminated by rapid cooling to 4°C, and after centrifugation, the supernatant was subjected to LC-MS analysis.
3. Pure Dibenzylfluorescein, fluorescein benzyl ester, fluorescein benzyl ether, and fluorescein (all at 10 μM) were analyzed as standards in the absence and presence of 2 M NaOH.
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Chemical Information
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CAS No. 97744-44-0
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Appearance Solid
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Molecular Weight 512.55
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Formula C34H24O5
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Color Yellow to orange
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SMILES
O=C1C=C2OC3=CC(OCC4=CC=CC=C4)=CC=C3C(C5=CC=CC=C5C(OCC6=CC=CC=C6)=O)=C2C=C1
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Synonyms
DBF
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
Solvent & Solubility
DMSO : 50 mg/mL (97.55 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Purity & Documentation
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Data Sheet (275 KB)
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SDS (393 KB)
- English - EN (393 KB)
- Français - FR (393 KB)
- Deutsch - DE (393 KB)
- Norwegian - NO (393 KB)
- Español - ES (393 KB)
- Swedish - SV (393 KB)
- Italian - IT (393 KB)
- Portuguese - PT (393 KB)
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Handling Instructions (2659 KB)
References
[1]. Stresser DM., et al. Substrate-dependent modulation of CYP3A4 catalytic activity: Analysis of 27 test compounds with four fluorometric substrates. Drug Metabolism and Disposition 28(12), 1440-1448 (2000). [Content Brief]
[2]. Donato MT., et al. Fluorescence-based assays for screening nine cytochrome P450 (P450) activities in intact cells expressing individual human P450 enzymes. Drug Metab. Dispos. 32(7), 699-706 (2004). [Content Brief]
[3]. Salminen KA, et al. Simple, direct, and informative method for the assessment of CYP2C19 enzyme inactivation kinetics. Drug Metabolism and Disposition 39(3), 412-418 (2011). [Content Brief]
[4]. Moutinho D, et al. Altered human CYP3A4 activity caused by Antley-Bixler syndrome-related variants of NADPH-cytochrome P450 oxidoreductase measured in a robust in vitro system. Drug Metabolism and Disposition 40(4), 754-760 (2012). [Content Brief]
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 1.9510 mL | 9.7551 mL | 19.5103 mL | 48.7757 mL |
| 5 mM | 0.3902 mL | 1.9510 mL | 3.9021 mL | 9.7551 mL | |
| 10 mM | 0.1951 mL | 0.9755 mL | 1.9510 mL | 4.8776 mL | |
| 15 mM | 0.1301 mL | 0.6503 mL | 1.3007 mL | 3.2517 mL | |
| 20 mM | 0.0976 mL | 0.4878 mL | 0.9755 mL | 2.4388 mL | |
| 25 mM | 0.0780 mL | 0.3902 mL | 0.7804 mL | 1.9510 mL | |
| 30 mM | 0.0650 mL | 0.3252 mL | 0.6503 mL | 1.6259 mL | |
| 40 mM | 0.0488 mL | 0.2439 mL | 0.4878 mL | 1.2194 mL | |
| 50 mM | 0.0390 mL | 0.1951 mL | 0.3902 mL | 0.9755 mL | |
| 60 mM | 0.0325 mL | 0.1626 mL | 0.3252 mL | 0.8129 mL | |
| 80 mM | 0.0244 mL | 0.1219 mL | 0.2439 mL | 0.6097 mL |
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