DAF-2 diacetate

DAF-2 diacetate (CAS No.: 205391-02-2) is a widely used cell-permeable fluorescent probe for the sensitive detection and bioimaging of nitric oxide (NO), a key signaling molecule involved in diverse physiological and pathological processes. As a diacetate derivative of DAF-2, it readily diffuses across cellular membranes and is subsequently hydrolyzed by intracellular esterases to yield the less permeable active form, which is retained within cells and enables stable signal accumulation. In the presence of oxygen, the probe reacts with NO to form a highly fluorescent triazole derivative, allowing quantitative monitoring of NO production with high sensitivity, typically in the nanomolar range under physiological conditions. This mechanism makes it particularly suitable for studying NO-related signaling pathways, including those associated with endothelial function, neuronal communication, and immune responses, where NO acts as a mediator of vasodilation, neurotransmission, and inflammatory regulation. DAF-2 diacetate is broadly applied in live-cell imaging, in vitro biochemical assays, and in vivo models to investigate NO dynamics in both low-output systems such as endothelial cells and high-output systems such as activated macrophages. Its robust fluorescence response and compatibility with standard excitation and emission settings support its use in high-content screening and drug discovery workflows aimed at modulating NO-associated pathways, with experimental concentrations typically optimized according to specific cellular systems and assay conditions.