1. Biochemical Assay Reagents

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

Cat. No.: HY-D0719 Purity: 99.56%
Handling Instructions

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.
Fluorescein Diacetate Chemical Structure

Fluorescein Diacetate Chemical Structure

CAS No. : 596-09-8

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Free Sample (0.5-1 mg)   Apply now  
10 mM * 1 mL in DMSO USD 66 In-stock
5 g USD 60 In-stock
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Customer Review

  • Biological Activity

  • Protocol

  • Technical Information

  • Purity & Documentation

  • References


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].

Clinical Trial
Preparing Stock Solutions
Concentration Volume 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
10 mM 0.2402 mL 1.2008 mL 2.4017 mL
Please refer to the solubility information to select the appropriate solvent.
Cell Assay

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.

Molecular Weight









4°C, protect from light


Room temperature in continental US; may vary elsewhere

Solvent & Solubility

DMSO: ≥ 48 mg/mL

* "<1 mg/mL" means slightly soluble or insoluble. "≥" means soluble, but saturation unknown.

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