Dibenzylfluorescein  (Synonyms: DBF)

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 K_m value of 0.87-1.9 µM (Ex=485nm，Em=535nm). Dibenzylfluorescein is used to detect changes in CYP catalytic activity caused by drugs or disease^[1][2][3][4].

P450 催化的 Dibenzylfluorescein 代谢和碱的影响实验方法^[3]：
反应过程：Dibenzylfluorescein 被 P450 脱烷基生成荧光素苄基酯，如果存在氢氧化钠可使其水解成荧光素。加入 2 M 氢氧化钠也会使 Dibenzylfluorescein 分解成荧光素苄基醚。
1. CYP2C19 酶活性和抑制实验条件 150 μL 反应体系为： 0.1 M Tris-HCl 缓冲液 (pH 7.4)、10 μM Dibenzylfluorescein、15 pmol CYP2C19 酶和 NADPH 再生系统 50 μL。NADPH 再生系统体系为：1.13 mM NADP、12.5 mM 异柠檬酸、56.33 mM KCl、187.5 mM Tris-HCl、pH 7.4、12.5 mM MgCl2、0.0125 mM MnCl2 和 0.075 U/ml 异柠檬酸脱氢酶。
2.样品在 37°C 下孵育 30-60 分钟。快速冷却至 4°C 终止反应，离心后上清液进行 LC-MS 分析。
3. 在没有和有 2 M NaOH 情况下，以纯 Dibenzylfluorescein、荧光素苄酯、荧光素苄醚和荧光素 (均为 10 μM) 为标准品进行分析。

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

512.55

C₃₄H₂₄O₅

97744-44-0

固体

Yellow to orange

Room temperature in continental US; may vary elsewhere.

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

* 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (protect from light)。-80°C储存时，请在6个月内使用，-20°C储存时，请在1个月内使用。

• [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]