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  4. Fluorescein Diacetate

Fluorescein Diacetate  (Synonyms: 3,6-Diacetoxyfluoran; Di-O-acetylfluorescein)

Cat. No.: HY-D0719 Purity: 99.79%
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Fluorescein diacetate is a cell permeable esterase-substrate. Fluorescein diacetate can be used as a fluorogenic substrate for hGSTP1-1.

For research use only. We do not sell to patients.

CAS No. : 596-09-8

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Based on 10 publication(s) in Google Scholar

Other Forms of Fluorescein Diacetate:

Fluorescein Diacetate Related Antibodies

Top Publications Citing Use of Products

    Fluorescein Diacetate purchased from MedChemExpress. Usage Cited in: Adv Mater. 2025 Apr 29:e2418560.  [Abstract]

    The Fluorescein Diacetate (FDA; 100 μM; 20 min) staining images and brightfield (BF) images of SH-SY5Y cells, with validation of neuronal axonal growth repair by RMC/pApoE2 after co-culturing.

    Fluorescein Diacetate purchased from MedChemExpress. Usage Cited in: Collagen Leather. 2025 Nov 19，7(1):39.

    Representative images (CLSM) of Fluorescein Diacetate (FDA)/PI staining of MSCs in different hydrogels on day 1, 3, and 7 in vitro culture.

    Fluorescein Diacetate purchased from MedChemExpress. Usage Cited in: Toxicol Appl Pharmacol. 2025 Sep:502:117454.

    Fluorescence images of the inhibitory effect of Carbofuran on CES2 in HepG2 cells with Fluorescein Diacetate (FD; 10 μM; 30 min). (a) 0 μM; (b) 5 μM; (c) 50 μM; (d) 100 μM;

    Fluorescein Diacetate purchased from MedChemExpress. Usage Cited in: Cell Prolif. 2024 Apr 16:e13646.  [Abstract]

    Bright-field and Fluorescein Diacetate (FDA) fluorescence (green) images of single MCF-7 (top) and CT-26 (bottom) cells before and 2 h after the first sampling. Yellow arrows indicate the same cell before and after sampling; black shadows denote the nanocapillary.

    Fluorescein Diacetate purchased from MedChemExpress. Usage Cited in: Cell Death Discov. 2021 Dec 4;7(1):374.  [Abstract]

    Live cells and dead HSC-LX2 cells were assayed by Fluorescein Diacetate (FDA) staining and PI staining, respectively.

    • Biological Activity

    • Protocol

    • Purity & Documentation

    • References

    • Customer Review

    Description

    Fluorescein diacetate is a cell permeable esterase-substrate. Fluorescein diacetate can be used as a fluorogenic substrate for hGSTP1-1.

    In Vitro

    Fluorescein diacetate (FDA) is an acetylated derivative of the green fluorescent dye fluorescein[1]. Fluorescein diacetate (FDA), a fluorescent probe used for vital staining, is a fluorescently activated by esterolytic activity of human Pi-class glutathione S-transferase (hGSTP1) selectively among various cytosolic GSTs. Fluorescence activation of Fluorescein diacetate susceptible to GST inhibitors is observed in MCF7 cells exogenously overexpressing hGSTP1, but not in cells overexpressing hGSTA1 or hGSTM1. Fluorescein diacetate can be used as a fluorogenic substrate for hGSTP1-1. To investigate whether the fluorescence activation is due to hGSTP1 activity, Fluorescein diacetate is incubated with recombinant hGSTP1-1 and GSH in vitro. Remarkable fluorescence activation is observed in the presence of both hGSTP1-1 and GSH, whereas only slight activation is observed in the absence of either of them or when the enzyme is heat inactivated. This suggests that the fluorescence activation of Fluorescein diacetate depends on hGSTP1-1 activity. From the linear relationship between the rate of increase in fluorescence and the hGSTP1-1 concentration, the specific activity of the enzyme for 1 μM Fluorescein diacetate is determined to be 79±15 nmol/min/mg protein. Fluorescein diacetate is applicable as a fluorogenic substrate for evaluating inhibitors of GSTP1-1 in vitro. For Fluorescein diacetate as a substrate, both Ethacrynic acid (EA) and NBDHEX suppress the hGSTP1-1-dependent fluorescent increase in a concentration-dependent manner, with IC50s of 3.3±0.5 μM and 0.61±0.04 μM, respectively[2].

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

    Fluorescein Diacetate Related Antibodies
    Molecular Weight

    416.38

    Formula

    C24H16O7
    CAS No.
    Appearance

    Solid

    Color

    Light yellow to yellow

    SMILES

    O=C1OC2(C3=C(OC4=C2C=CC(OC(C)=O)=C4)C=C(OC(C)=O)C=C3)C5=C1C=CC=C5
    Shipping

    Room temperature in continental US; may vary elsewhere.
    Storage

    -20°C, protect from light

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

    Solvent & Solubility
    In Vitro: 

    DMSO : 23.33 mg/mL (56.03 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 2.4017 mL 12.0083 mL 24.0165 mL
    5 mM 0.4803 mL 2.4017 mL 4.8033 mL
    View the 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.

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    Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

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    Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

    This equation is commonly abbreviated as: C1V1 = C2V2

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    In Vivo Dissolution Calculator
    Please enter the basic information of animal experiments:

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    Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
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    Working solution concentration: mg/mL
    Purity & Documentation

    Purity: 99.79%

    SDS (393 KB)

    COA (252 KB) HNMR (269 KB) LCMS (93 KB)

    Handling Instructions (2659 KB)
    References
    Cell Assay
    [2]

    MCF7 cells (2-3×105) are seeded in a 35-mm glass bottom dish before the experiment. Prior to imaging, cells are washed with 1 mL PBS, then incubated in 1 mL Hanks’ Balanced Salt solution (HBSS(+) without phenol red) containing 1 μM Fluorescein diacetate (0.1% DMSO as a cosolvent) for 5 min at 37°C. Cells are washed twice with 1 mL PBS and 1 mL HBSS is added before imaging. Fluorescence images of fluorescein and DsRed-Express2 were acquired in the FITC channel (excitation at 473 nm) and the DsRed channel (excitation 559 nm). The 16-bit images obtained are analysed[2].

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

    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 2.4017 mL 12.0083 mL 24.0165 mL 60.0413 mL
    5 mM 0.4803 mL 2.4017 mL 4.8033 mL 12.0083 mL
    10 mM 0.2402 mL 1.2008 mL 2.4017 mL 6.0041 mL
    15 mM 0.1601 mL 0.8006 mL 1.6011 mL 4.0028 mL
    20 mM 0.1201 mL 0.6004 mL 1.2008 mL 3.0021 mL
    25 mM 0.0961 mL 0.4803 mL 0.9607 mL 2.4017 mL
    30 mM 0.0801 mL 0.4003 mL 0.8006 mL 2.0014 mL
    40 mM 0.0600 mL 0.3002 mL 0.6004 mL 1.5010 mL
    50 mM 0.0480 mL 0.2402 mL 0.4803 mL 1.2008 mL
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    Fluorescein Diacetate Related Classifications

    Help & FAQs
    • Do most proteins show cross-species activity?

      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

    Keywords:

    Fluorescein Diacetate596-09-83,6-Diacetoxyfluoran Di-O-acetylfluoresceinFluorescent DyeInhibitorinhibitorinhibit

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