Fluorescein diacetate

SKU: orb1307068

Description

Fluorescein diacetate (Di-O-acetylfluorescein) is a cell-permeable, non-fluorescent probe that is hydrolyzed by esterase activity, notably by human glutathione S-transferase P1-1 (hGSTP1-1), to yield fluorescent fluorescein. It is widely used in both in vitro and in vivo research for monitoring cellular esterase/GST activity, cell viability, and proliferation assays.

Key Properties

• CAS Number: 596-09-8
• MW: 416.38
• Purity: 99.79% (May vary between batches)
• Formula: C₂₄H₁₆O₇
• SMILES: CC(=O)Oc1ccc2c(Oc3cc(OC(C)=O)ccc3C22OC(=O)c3ccccc23)c1
• Target: Others
• Solubility: DMSO:6.88 mg/mL (16.52 mM)

Bioactivity

• In Vitro:
  Fluorescein diacetate (FDA) serves as a fluorogenic substrate in assessing the efficacy of inhibitors against the enzyme GSTP1-1 in vitro. Notably, Ethacrynic acid (EA) and NBDHEX are effective in inhibiting the enzyme-activated fluorescence of FDA, demonstrating concentration-dependent inhibition with IC50 values of 3.3±0.5 μM and 0.61±0.04 μM, respectively. FDA, an acetylated variant of fluorescein, is activated fluorescently through the esterolytic action of the human Pi-class glutathione S-transferase (hGSTP1), marking it specific among various cytosolic GSTs. This fluorogenic activity allows for the usage of FDA in vital staining. Particularly, when MCF7 cells overexpress hGSTP1, an observable fluorescence activation due to GST inhibitors is noted, an effect not seen in cells overexpressing hGSTA1 or hGSTM1. The enzyme-dependent fluorescence activation is further corroborated by incubating FDA with recombinant hGSTP1-1 and GSH, showing significant fluorescence only in the presence of both, indicating the necessity of hGSTP1-1 activity for such activation. This specificity is underscored by the established linear relation between fluorescence increase rate and hGSTP1-1 concentration, determining the enzyme's specific activity for FDA to be 79±15 nmol/min/mg protein.
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Storage & Handling

• Storage: -20°C
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

Fluorescein, Fluorescein diacetate, Fluorescein Diacetate, inhibit, Inhibitor, Di-O-acetylfluorescein, 3,6-Diacetoxyfluoran

Compound Identifiers

PubChem CID

65047