DAF-FM DA
Based on 17 publication(s) in Google Scholar
DAF-FM DA (Diaminofluorescein-FM diacetate) is a fluorescent probe for the detection and bioimaging of nitric oxide (NO). DAF-FM DA spontaneously crosses the plasma membrane and is subsequently cleaved by esterases to generate intracellular DAF-FM (Ex/Em=495/515 nm).
For research use only. We do not sell to patients.
- Purity: 98.32%
- CAS No.: 254109-22-3
- Formula: C25H18F2N2O7
- Molecular Weight:496.42
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Storage:
-20°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Purity & Documentation
- References
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Help & FAQs
Publications Citing Use of MedChemExpress (MCE) DAF-FM DA
More- Bioact Mater. 2022 Mar 17;18:91-103. [Abstract]
- Redox Biol. 2023 Dec:68:102952. [Abstract]
- Biomaterials. 2026 Aug:331:124126. [Abstract]
- Chem Eng J. 2023 Dec 2, 147850.
- J Hazard Mater. 2024 Jul 5:472:134485. [Abstract]
- Regen Biomater. 2026 May 5:13:rbag086. [Abstract]
- J Invest Dermatol. 2025 Sep 15:S0022-202X(25)02428-5. [Abstract]
- Life Sci. 2026 Sep 15:401:124502. [Abstract]
- ACS Biomater Sci Eng. 2023 May 8;9(5):2545-2557. [Abstract]
- Eur J Pharmacol. 2025 Sep 5:1002:177807. [Abstract]
- Int Immunopharmacol. 2026 Feb 1:170:116055. [Abstract]
- Int Immunopharmacol. 2024 Oct 31;143(Pt 3):113433. [Abstract]
- Food Safety and Health. 2026 Jan 7.
- Biochim Biophys Acta Mol Basis Dis. 2024 Apr;1870(4):167110. [Abstract]
- Exp Cell Res. 2024 Mar 20;437(1):113998. [Abstract]
- Exp Parasitol. 2020 Aug:215:107917. [Abstract]
- bioRxiv. 2025 Jun 27:2025.06.24.660441. [Abstract]
Customer Validation & Images
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IF
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Cell Imaging/Staining
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Cell Imaging/Staining
Biological Activity
Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
1.1 Preparation of stock solution[2]
Use anhydrous DMSO to prepare a 5 mM DAF-FM DA stock solution.
Note: It is recommended that the DAF-FM DA stock solution be stored at -20°C or -80°C in the dark after aliquoting.
1.2 Preparation of working solution
Dilute the stock solution with Hank’s buffer or serum-free medium to prepare a 5 μM DAF-FM DA working solution.
Note: Please adjust the concentration of DAF-FM DA working solution according to actual conditions and prepare it before use.
2 Staining steps
1) Seed breast cancer cells in a 6-well plate.
2) Aspirate the culture medium.
3) Add 5 μM DAF-FM DA working solution to each well and incubate at 37°C in the dark for 30 min to allow the probe to enter the cells.
4) Replace with fresh Hank’s buffer and continue incubation for 20 min to complete the deesterification reaction.
5) Counterstain the cell nucleus with Hoechst dye.
6) Quantify the fluorescence intensity using a microplate reader at 488 nm excitation and 515 nm emission, and normalize to the number of cells stained with Hoechst.
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
515
495
Chemical Information
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CAS No. 254109-22-3
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Appearance Solid
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Molecular Weight 496.42
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Formula C25H18F2N2O7
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Color Off-white to yellow
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SMILES
O=C1OC2(C3=C(OC4=C2C=C(F)C(OC(C)=O)=C4)C=C(OC(C)=O)C(F)=C3)C5=C1C(N)=C(NC)C=C5
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Synonyms
Diaminofluorescein-FM diacetate
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
-20°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
Publications (17)
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Journal Impact Factor
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Most Recent
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Bioact Mater
Upconversion nanoparticles regulated drug & gas dual-effective nanoplatform for the targeting cooperated therapy of thrombus and anticoagulation. [Abstract]2022 Mar 17;18:91-103. PMID: 35387173
DAF-FM DA purchased from MedChemExpress. Usage Cited in: Bioact Mater. 2022 Mar 17;18:91-103. [Abstract]
The fluorescent images of HUVECs stained with NO fluorescent probe (DAF-FM DA, 5 μMat 37 °C for 1 h) and Hoechst 33342 after being treated with UCATS under dark conditions or 808 nm NIR irradiation (1 W cm−2).
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Redox Biol
RelB-activated GPX4 inhibits ferroptosis and confers tamoxifen resistance in breast cancer. [Abstract]2023 Dec:68:102952. PMID: 37944384
DAF-FM DA purchased from MedChemExpress. Usage Cited in: Redox Biol. 2023 Dec:68:102952. [Abstract]
The levels of intercellular nitric oxide (NO) in TAM-treated cells were analyzed using DAF-FM DA (5 μM, 30 min) with nuclear staining.
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Biomaterials
Conjugated polymer intrachain donor-acceptor reconfiguration-tailored off-on NIR-II photoacoustic probes with ultralow background for high-fidelity imaging of immunotherapy-associated macrophage reprogramming. [Abstract]2026 Aug:331:124126. PMID: 41830766 -
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J Hazard Mater
Effect of oxidative stress induced by 2,3,7,8- tetrachlorodibenzo-p-dioxin on DNA damage. [Abstract]2024 Jul 5:472:134485. PMID: 38701725 -
Regen Biomater
A 'frost formation'-inspired near-infrared-responsive nitric oxide-releasing hydrogel for enhancing fat graft survival. [Abstract]2026 May 5:13:rbag086. PMID: 42206192 -
J Invest Dermatol
Inducible Nitric Oxide Deficiency Attenuates Necroptosis in Psoriasis-like Dermatitis in Mice through Reducing Oxidative Stress. [Abstract]2025 Sep 15:S0022-202X(25)02428-5. PMID: 40962201 -
Life Sci
Nitroxyl relieves acute kidney injury by suppressing SLC31A1-mediated cuproptosis in renal tubular epithelial cells. [Abstract]2026 Sep 15:401:124502. PMID: 42214610 -
ACS Biomater Sci Eng
Biomimetic Biomembrane Encapsulation and Targeted Delivery of a Nitric Oxide Release Platform for Therapy of Parkinson's Disease. [Abstract]2023 May 8;9(5):2545-2557. PMID: 37040524 -
Eur J Pharmacol
Morin ameliorates coronary artery relaxation by activating TRPV4-eNOS-NO signalling in high-salt diet-fed rats. [Abstract]2025 Sep 5:1002:177807. PMID: 40472987 -
Int Immunopharmacol
Targeting periodontal inflammatory microenvironment to ameliorate periodontitis: A nitrogen ions implantation technique. [Abstract]2026 Feb 1:170:116055. PMID: 41448003
DAF-FM DA purchased from MedChemExpress. Usage Cited in: Int Immunopharmacol. 2026 Feb 1:170:116055. [Abstract]
The NO levels in the gingival tissues of mice were detected by IF assay through the DAF-FM DA staining at 4, 24 and 48 h after exposure to 2-min PBNI or 20 mM L-Arg. The pink rectangles in low-power-field images highlight the picture ROI and orange dotted lines indicate the boundary of epithelium and lamina propria.
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Int Immunopharmacol
Interleukin-33 induces angiogenesis after myocardial infarction via AKT/eNOS signaling pathway. [Abstract]2024 Oct 31;143(Pt 3):113433. PMID: 39486188 -
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Biochim Biophys Acta Mol Basis Dis
Co-activation of Mas and pGCA receptors suppresses endothelin-1-induced endothelial dysfunction via nitric oxide/cGMP system. [Abstract]2024 Apr;1870(4):167110. PMID: 38462025 -
Exp Cell Res
Protein phosphatase 4 mediates palmitic acid-induced endothelial dysfunction by decreasing eNOS phosphorylation at serine 633 in HUVECs. [Abstract]2024 Mar 20;437(1):113998. PMID: 38513962 -
Exp Parasitol
Myeloid-derived suppressor cells exert immunosuppressive function on the T helper 2 in mice infected with Echinococcus granulosus. [Abstract]2020 Aug:215:107917. PMID: 32446699 -
bioRxiv
Flow-sensitive HEG1 controls eNOS activity to prevent endothelial dysfunction, hypertension, and atherosclerosis. [Abstract]2025 Jun 27:2025.06.24.660441. PMID: 40667131
Solvent & Solubility
DMSO : 2.5 mg/mL (5.04 mM; Need ultrasonic and warming; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Purity & Documentation
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Data Sheet (280 KB)
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SDS (393 KB)
- English - EN (393 KB)
- Français - FR (393 KB)
- Deutsch - DE (393 KB)
- Norwegian - NO (393 KB)
- Español - ES (393 KB)
- Swedish - SV (393 KB)
- Italian - IT (393 KB)
- Korean - KR (393 KB)
- Portuguese - PT (393 KB)
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Handling Instructions (2659 KB)
References
[1]. Ann Huysseune, et al. Bone Formation in Zebrafish: The Significance of DAF-FM DA Staining for Nitric Oxide Detection. Biomolecules. 2023 Dec 12;13(12):1780. [Content Brief]
[2]. Zhi Xu, et al. RelB-activated GPX4 inhibits ferroptosis and confers tamoxifen resistance in breast cancer. Redox Biol. 2023 Dec:68:102952. [Content Brief]
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 2.0144 mL | 10.0721 mL | 20.1442 mL | 50.3606 mL |
| 5 mM | 0.4029 mL | 2.0144 mL | 4.0288 mL | 10.0721 mL |
DAF-FM DA Related Classifications
HY-D0717 Related Classifications
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