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| Product Name | DAF-FM DA Nitric Oxide Fluorescent Probe |
| Catalog No. | RESK-0560 |
| Description | DAF-FM DA (4-amino-5-methylamino-2',7'-difluorofluorescein diacetate, also known as DAF-FM diacetate) is a next-generation fluorescent probe for the quantitative detection of nitric oxide (NO). The cell-permeant diacetate crosses the membrane of live cells and is hydrolyzed by intracellular esterases to membrane-impermeant DAF-FM. DAF-FM itself shows only very weak fluorescence (quantum yield ~0.005), but upon reaction with NO it forms a benzotriazole derivative that emits strong green fluorescence (quantum yield ~0.81) with excitation/emission maxima of approximately 495/515 nm. Compared with the earlier widely used NO probe DAF-2 diacetate, DAF-FM DA offers three improvements: the fluorescence of its NO adduct is pH-independent above pH 5.5; the reaction product is more photostable and less prone to quenching, facilitating detection; and the sensitivity is higher, with a minimum detectable NO concentration of 3 nM versus approximately 5 nM for DAF-2 diacetate. |
| Intended Use | For quantitative detection of intracellular nitric oxide (NO) levels in live cells. Recommended working concentration: 1-10 μM. If cells are harvested before probe loading, one package typically supports at least 100 samples. |
| Principle / Technology | Cell-permeant DAF-FM DA enters live cells and is converted by cytosolic esterases into DAF-FM, which cannot cross the cell membrane. DAF-FM reacts with NO to form a highly fluorescent benzotriazole product, and the resulting green fluorescence intensity is proportional to the NO level. |
| Detection Method | Fluorescence detection (Ex ~495 nm, Em ~515 nm). Any instrument capable of detecting fluorescein can be used, including fluorescence microscopes, laser scanning confocal microscopes, flow cytometers, fluorescence spectrophotometers, and fluorescence microplate readers. |
| Sample Type | Live cells (intracellular NO detection). |
| Performance Range / Specifications | Appearance: white powder CAS No.: 254109-22-3 Molecular formula: C25H18F2N2O7 Molecular weight: 496.42 Purity: >98% Ex/Em: ~495/515 nm Quantum yield: ~0.005 (free probe) to ~0.81 (NO adduct) |
| Sensitivity / LOD | Minimum detectable NO concentration: 3 nM (versus approximately 5 nM for DAF-2 diacetate). |
| Specificity | Reacts with nitric oxide to form a fluorescent benzotriazole adduct; the fluorescence of the NO adduct is independent of pH above pH 5.5. |
| Reaction Conditions / Protocol | Prepare a stock solution in DMSO, then dilute with PBS to obtain the working solution. Recommended working concentration: 1-10 μM. Load the probe into live cells and measure the green fluorescence signal. |
| Components / Formulation | Single-component reagent supplied as a white powder. |
| Storage Conditions | Store at -25 to -15 °C. |
| Shelf Life | 2 years when stored at -25 to -15 °C. |
| Package Specifications | 50 μg; 10× 50 μg |
| Product Form | White powder. |
| Purity | >98% |
| Concentration | Recommended working concentration: 1-10 μM. |
| Molecular Weight | 496.42 |
| pH Range / Optimal pH | The fluorescence of the NO adduct is pH-independent above pH 5.5. |
| Expiration Date / Stability | Stable for 2 years at -25 to -15 °C. The NO reaction product is more photostable and less susceptible to quenching than that of earlier-generation NO probes. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Compatible with fluorescence microscopes, laser scanning confocal microscopes, flow cytometers, fluorescence spectrophotometers, and fluorescence microplate readers. |
| Recommended Buffer System | DMSO for stock solution preparation; PBS for working solution dilution. |
| Application Notes / Precautions | Dissolve in DMSO to prepare a stock solution and dilute into PBS to prepare the working solution before use. |
For research use only, not for clinical use.
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