NBD-Pen
Based on 1 Customer Validation
NBD-Pen is the first fluorescence probe for lipid radicals with high selectivity and sensitivity (λex: 470 nm, λem: 530 nm). NBD-Pen specifically detects lipid derived radicals over other reactive species present in biological systems, including H2O2, ClO-, O2-?, and ?OH. NBD-Pen directly detects lipid radicals in living cells by turn-on fluorescence. NBD-Pen decreases inflammation, apoptosis, and oxidative stress markers. NBD-Pen can be studied in various disease models such as hepatic carcinoma.
For research use only. We do not sell to patients.
- Purity: 99.94%
- CAS No.: 1955505-54-0
- Formula: C19H28N5O4*
- Molecular Weight:390.46
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Storage:
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
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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
Biological Activity
Guide (The following is our recommend protocol. This protocol is only a guide and should be modified according to your specific needs).
1. Preparation of stock solution
Use anhydrous DMSO to prepare a 10 mM NBD-Pen stock solution.
2. Preparation of working solution
Use preheated serum-free cell culture medium or PBS to dilute the stock solution to prepare a 10 μM NBD-Pen working solution.
Note: Please adjust the concentration of NBD-Pen working solution according to actual conditions and prepare it before use.
3. Spectroscopic methods
3.1 Use 100 mM phosphate buffer and 0.5% ethanol to prepare a lipid (500 μM) emulsion.
3.2 Mix this emulsion with 5 μM NBD-Pen in phosphate buffer containing 0.5% acetonitrile.
3.3 Add the LOX into the mixture at indicated concentration in phosphate buffer and incubate.
3.4 Perform fluorescence emission and ESR measurements.
3.5 Measure fluorescence spectra of the probes at excitation and emission wavelengths of 470 and 530 nm.
3.6 Monitor ESR spectrometer using a X-band (9.45 GHz) ESR spectrometer.
4. Preparation and staining of cells
4.1 Discard cell culture medium and rinse cells three times with PBS and phenol red free culture media with 10% FBS.
4.2 Add 1 μM of NBD-Pen and incubate for 10 min.
4.3 Add 30 mM of Diethylnitrosamine (DEN).
4.4 Perform fluorescence imaging immediately at 37°C under a humidified atmosphere of 5% CO2 in air.
4.5 Administer Hoechst33342 (1 μM) and SKF525A (50 μM) and incubate for 1 h, then wash with PBS.
4.6 Conduct fluorescence imaging with a confocal laser-scanning microscope with 63x or 40x objective lens.
4.7 Use the following detection lasers: Hoechst33342, λex = 405 nm, λem = 410-505 nm; NBD-Pen, λex = 458 nm, λem = 490-674 nm.
NBD-Pen (1 μM, 20 min) shows low background fluorescence in Hepal-6 cells[1].
NBD-Pen (1 μM, 10 min, followed by 30 mM DEN for another 20 min) enables visualization of intracellular lipid radicals produced by DEN activation in Hepa1-6 cells[1].
NBD-Pen (10 μM, 0-60 min) induces increased fluorescent intensity in a concentration-dependent manner in LDL with the addition of hemin (HY-19424) (0-3μM)[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
NBD-Pen (2 mM, i.v., 1-24 h after a 1 g/kg iron dectran administration) significantly increases the fluorescence intensity in LDL at 1 h after iron administration to mice with the peak at 2 h in iron overloaded model mice[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:Rats pre-treated with SKF-525A (i.p.)[1]
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Dosage:2 mM
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Administration:Intravenous injection (i.v.) 1 h after DEN administrations
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Result:Indicated that NBD-Pen had favorable properties, including stability, a turn-on switching function and the ability to detect lipid radicals in living animals.
Chemical Information
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CAS No. 1955505-54-0
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Appearance Solid
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Molecular Weight 390.46
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Formula C19H28N5O4*
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Color Orange to red
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SMILES
[O]N1C(C)(C)CC(NC2=CC=C([N+]([O-])=O)C3=NON=C32)CC1(C)CCCCC
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
Solvent & Solubility
DMSO : 100 mg/mL (256.11 mM; Need ultrasonic; 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 (277 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Handling Instructions (2659 KB)
References
[1]. Yamada, K., et al., (2016). Fluorescence probes to detect lipid-derived radicals. Nature chemical biology, 12(8), 608–613. [Content Brief]
[2]. Ishida Y, et al. Detection and inhibition of lipid-derived radicals in low-density lipoprotein. Free Radic Biol Med. 2017 Dec;113:487-493. [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 | 2.5611 mL | 12.8054 mL | 25.6108 mL | 64.0270 mL |
| 5 mM | 0.5122 mL | 2.5611 mL | 5.1222 mL | 12.8054 mL | |
| 10 mM | 0.2561 mL | 1.2805 mL | 2.5611 mL | 6.4027 mL | |
| 15 mM | 0.1707 mL | 0.8537 mL | 1.7074 mL | 4.2685 mL | |
| 20 mM | 0.1281 mL | 0.6403 mL | 1.2805 mL | 3.2014 mL | |
| 25 mM | 0.1024 mL | 0.5122 mL | 1.0244 mL | 2.5611 mL | |
| 30 mM | 0.0854 mL | 0.4268 mL | 0.8537 mL | 2.1342 mL | |
| 40 mM | 0.0640 mL | 0.3201 mL | 0.6403 mL | 1.6007 mL | |
| 50 mM | 0.0512 mL | 0.2561 mL | 0.5122 mL | 1.2805 mL | |
| 60 mM | 0.0427 mL | 0.2134 mL | 0.4268 mL | 1.0671 mL | |
| 80 mM | 0.0320 mL | 0.1601 mL | 0.3201 mL | 0.8003 mL | |
| 100 mM | 0.0256 mL | 0.1281 mL | 0.2561 mL | 0.6403 mL |
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