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Fluorescein Diacetate

CAS No. : 596-09-8

MCE 站：Fluorescein Diacetate

产品活性：Fluorescein diacetate 是一种细胞渗透性的酯酶底物。Fluorescein diacetate 可作为人谷胱甘肽硫转移酶 Pi (hGSTP1-1) 的荧光底物。

研究领域：Others

作用靶点：Others

In Vitro: Fluorescein diacetate (FDA) is an acetylated derivative of the green fluorescent dye fluorescein. 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.

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