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| Product Name | 3D Cell Nitric Oxide Detection Kit (DAF-FM DA), 200 Tests |
| Catalog No. | REKI-0001 |
| Description | This kit detects nitric oxide in three-dimensionally cultured spheroids and organoids quickly and with high sensitivity. Its DAF-FM DA probe improves on the older DAF-2 diacetate in three documented ways: the fluorescent adduct it forms with NO is insensitive to pH above 5.5, the product's fluorescence is more stable and resists quenching, and the detection floor reaches 3 nM against about 5 nM for the older probe. A 20-minute stain is enough to produce bright green fluorescence inside cell spheres and organoids under the microscope. |
| Intended Use | Rapid, sensitive fluorescence measurement of nitric oxide in 3D-cultured cell spheroids and organoids. |
| Principle / Technology | DAF-FM DA crosses the live cell membrane, where cytoplasmic esterases convert it to membrane-impermeant DAF-FM. Weakly fluorescent on its own (quantum yield about 0.005), DAF-FM reacts with NO to form a fluorescent benzotriazole with a quantum yield of about 0.81, giving strong green emission. |
| Detection Method | Green fluorescence at Ex 495 nm / Em 515 nm; any fluorescein-capable instrument - fluorescence microscope, confocal microscope, flow cytometer, fluorescence spectrophotometer or plate reader. |
| Sample Type | 3D-cultured cell spheroids or organoids (live). |
| Performance Range / Specifications | Detection floor 3 nM; staining time 20 minutes; quantum yield rises from about 0.005 before the NO reaction to about 0.81 after; Ex 495 nm / Em 515 nm; pH-independent above pH 5.5; kit size 200 tests. |
| Sensitivity / LOD | Detection limit 3 nM (compared with about 5 nM for the previous-generation DAF-2 diacetate). |
| Specificity | Selective for nitric oxide; fluorescent product is stable and pH-independent above pH 5.5. |
| Reaction Conditions / Protocol | Load the spheroids or organoids with the DAF-FM DA working solution, incubate for 20 minutes protected from light, wash as required, then record the green fluorescence on the chosen instrument. |
| Components / Formulation | DAF-FM DA (1000×) and NOup (1000×), both stored protected from light; kit size 200 tests. |
| Storage Conditions | Store at -25 to -15 °C, protected from light, for 12 months; keep both the DAF-FM DA (1000×) and NOup (1000×) components protected from light. |
| Shelf Life | 12 months under the recommended storage conditions. |
| Package Specifications | 200 tests per kit |
| Product Form | Kit of probe and auxiliary reagent solutions. |
| Quality Control | Probe performance is anchored by the documented detection floor, quantum yield change and staining time. |
| Key Features | - Detection floor of 3 nM, beyond the previous-generation probe - Only 20 minutes of staining needed - Fluorescent product is pH-independent above pH 5.5 - Quantum yield jumps from about 0.005 to about 0.81 on reaction - Built for 3D spheroids and organoids, not just flat monolayers |
| Concentration | Components are supplied as 1000× concentrates. |
| Activity / Unit Definition | One kit provides 200 test reactions; performance is defined by the 3 nM detection floor. |
| Source / Origin | Synthetic DAF-FM DA fluorescent probe supplied with auxiliary reagent. |
| Expiration Date / Stability | Stable through the stated shelf life when kept protected from light at the recommended storage temperature. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Compatible with standard fluorescence instrumentation - microscopes, plate readers, flow cytometers or imaging systems as appropriate to the format. |
| Recommended Buffer System | The kit supplies concentrate-format reagents; working solutions are prepared by the user before staining. |
| Application Notes / Precautions | Protect both components from light at every step, keep the 20-minute staining window consistent, and confirm fluorescein-compatible filter sets before imaging 3D cultures, which scatter more light than flat monolayers. |
| Batch-to-Batch Consistency | Each production lot is released against fixed acceptance criteria, keeping performance consistent between orders. |
| Model Type | Fluorescent detection kit for 3D culture formats |
| Starting Material / Tissue Site | DAF-FM DA probe with NOup auxiliary reagent, supplied as 1000× concentrates |
| Delivery Standard | 200-test kit |
| Turnaround Time | No turnaround figure is published; shipping is arranged when the order is confirmed. |
| Readouts / Downstream Assays | Green fluorescence at Ex 495/Em 515 nm reporting nitric oxide. |
For research use only, not for clinical use.
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