RhoNox-4
Based on 1 Customer Validation
RhoNox-4 is a Fe2+-selective fluorescence-activatable probe and an inhibitor of ferroptosis. RhoNox-4 shows no obvious cytotoxicity even at concentrations up to 50 μM. RhoNox-4 can selectively oxidize labile Fe2+ to Fe3+, depleting the iron pool required for lipid peroxidation and ferroptosis. RhoNox-4 inhibits ferroptosis induced by ferroptosis inducers; its activity depends on localization to the endoplasmic reticulum/Golgi apparatus, and it does not affect apoptosis or necrosis. RhoNox-4 can be used for imaging labile Fe2+ in living cells, and also enables high-throughput screening of labile iron homeostasis regulators based on 96-well plates. RhoNox-4 allows visualization of cytoplasmic Fe2+ levels in root tips of *Arabidopsis* (Ex/Em = 530-560/573-647 nm).
For research use only. We do not sell to patients.
- Purity: 98.65%
- CAS No.: 2711762-70-6
- Formula: C38H44N4O8
- Molecular Weight:684.78
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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
Biological Activity
RhoNox-4 (2 μM; up to 3000 seconds) shows rapid, highly sensitive fluorescence turn-on response to Fe2+ in cell-free buffer, with >100-fold fluorescence enhancement within 15 minutes[1].
RhoNox-4 (1 μM) is a cell-permeable Fe2+-selective fluorescent probe that produces high-contrast fluorescence signals in Fe2+-treated HepG2 cells[1].
RhoNox-4 supports high-throughput screening for compounds modulating subcellular labile iron levels, identifying lomofungin as a novel ferritin-downregulating labile iron upregulator[1].
RhoNox-4 (3.2-12.8 μM) inhibits Erastin (HY-15763)-induced ferroptosis in HT1080 cells, maintaining ~90% cell viability at 3.2 μM and ~80% cell viability at 12.8 μM[1].
RhoNox-4 (0.2-12.8 μM; 12 h) potently inhibits Erastin-induced ferroptosis in HT1080 cells with an IC50 of approximately 3.5 μM[2].
RhoNox-4 (0.2-12.8 μM; 12 h) robustly inhibits RSL3 (HY-100218A)-induced ferroptosis in HT1080 cells, reducing cell death by more than 50%[2].
RhoNox-4 (12.8 μM; 12 h) inhibits FIN56 (HY-103087)- and FINO2 (HY-129457)-induced ferroptosis in HT1080 cells, providing effective cell viability rescue[2].
RhoNox-4 (12.8 μM; 12 h, 24 h) does not protect HT1080 cells from necrosis or apoptosis, demonstrating selective inhibition of ferroptosis[2].
RhoNox-4 (1.0 h, 6 h) detects elevated labile Fe2+ levels in erastin-treated HT1080 cells[2].
RhoNox-4 (2 μM; 1 h) acts as a highly selective, turn-on fluorescent probe for Fe2+ in cell-free conditions, with an off/on ratio of 160 and an apparent reaction rate of 1.0 × 10-3 s-1[3].
RhoNox-4 (1 μM; 30 min) efficiently detects labile Fe2+ in live HepG2 cells, with highest sensitivity in the endoplasmic reticulum and mitochondria, and shows negligible cytotoxicity up to 50 μM[3].
RhoNox-4 (5 µM; 3 h incubation) labels cytosolic Fe2+ in Arabidopsis thaliana root tips, revealing that Split +Fe root tips have higher Fe2+ content than Cont. +Fe root tips after 24 h of split-root culture treatment[4].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Cell Line:HT1080 human fibrosarcoma cells
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Concentration:0.2-12.8 μM (with 10 μM erastin)
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Incubation Time:12 h
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Result:Potently inhibited erastin-induced ferroptosis, with an IC50 value of 3.5 μM.
Significantly increased HT1080 cell viability compared to erastin-only treatment, achieving near-full rescue at higher concentrations.
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Cell Line:HT1080 human fibrosarcoma cells
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Concentration:0.2-12.8 μM (with 10 μM RSL3)
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Incubation Time:12 h
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Result:Demonstrated robust inhibitory activity against RSL3-induced ferroptosis, reducing cell death by more than 50%.
Significantly increased HT1080 cell viability compared to RSL3-only treatment.
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Cell Line:HT1080 human fibrosarcoma cells
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Concentration:12.8 μM (with lethal concentrations of FIN56 or FINO2)
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Incubation Time:12 h
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Result:Exhibited comparable inhibitory activity against FIN56- and FINO2-induced ferroptosis.
Effectively rescued HT1080 cell viability.
Chemical Information
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CAS No. 2711762-70-6
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Appearance Solid
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Molecular Weight 684.78
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Formula C38H44N4O8
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Color Off-white to pink
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SMILES
CC(C)(OC(N1CC[N+](C2=CC3=C(C4(C5=CC=C(C=C5O3)N6CCN(C(OC(C)(C)C)=O)CC6)OC(C7=C4C=CC=C7)=O)C=C2)(CC1)[O-])=O)C
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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)
Solvent & Solubility
DMSO : 50 mg/mL (73.02 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)
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 1.25 mg/mL (1.83 mM); Clear solution
This protocol yields a clear solution of ≥ 1.25 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (12.5 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 1.25 mg/mL (1.83 mM); Clear solution
This protocol yields a clear solution of ≥ 1.25 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (12.5 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL. * In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation
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Data Sheet (278 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]. Hirayama T, et al. -oxide-based and trioxolane-based fluorescent probes for monitoring labile iron and labile heme: an update. Journal of clinical biochemistry and nutrition. 2026 Jan;78(1):6-13. [Content Brief]
[2]. Kawai K, et al. Inhibition of ferroptosis by -oxide-based fluorescent probes selective oxidation of ferrous ions. Chemical science. 2025 Jun 25;16(25):11240-11245. [Content Brief]
[3]. Hirayama T, et al. High-Throughput Screening for the Discovery of Iron Homeostasis Modulators Using an Extremely Sensitive Fluorescent Probe. ACS sensors. 2020 Sep 25;5(9):2950-2958. [Content Brief]
[4]. Tabata R, et al. Systemic Regulation of Iron Acquisition by Arabidopsis in Environments with Heterogeneous Iron Distributions. Plant & cell physiology. 2022 Jun 15;63(6):842-854. [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 | 1.4603 mL | 7.3016 mL | 14.6032 mL | 36.5081 mL |
| 5 mM | 0.2921 mL | 1.4603 mL | 2.9206 mL | 7.3016 mL | |
| 10 mM | 0.1460 mL | 0.7302 mL | 1.4603 mL | 3.6508 mL | |
| 15 mM | 0.0974 mL | 0.4868 mL | 0.9735 mL | 2.4339 mL | |
| 20 mM | 0.0730 mL | 0.3651 mL | 0.7302 mL | 1.8254 mL | |
| 25 mM | 0.0584 mL | 0.2921 mL | 0.5841 mL | 1.4603 mL | |
| 30 mM | 0.0487 mL | 0.2434 mL | 0.4868 mL | 1.2169 mL | |
| 40 mM | 0.0365 mL | 0.1825 mL | 0.3651 mL | 0.9127 mL | |
| 50 mM | 0.0292 mL | 0.1460 mL | 0.2921 mL | 0.7302 mL | |
| 60 mM | 0.0243 mL | 0.1217 mL | 0.2434 mL | 0.6085 mL |
RhoNox-4 Related Classifications
HY-D3116 Related Classifications
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