| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
RhoNox-1 specifically targets labile ferrous iron (Fe2+) within live cells. Upon binding to Fe2+, it undergoes an irreversible chemical reaction, which converts the non-fluorescent or weakly fluorescent probe to a brightly fluorescent product, enabling visualization of Fe2+ distribution, particularly localized in the Golgi apparatus.
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| ln Vitro |
1. RhoNox-1 functioning solution components 1.1 To produce the stock solution, use 110 μL of anhydrous DMSO. To obtain a 1 mM stock solution, use 50 μg of RhoNox-1. 1.2 Elements of a functional solution To make a 1-10 μM RhoNox-1 working solution, use serum-free cell bone marrow or PBS storage solution that has already been made. Note: Please prepare RhoNox-1 working fluid for immediate usage by adjusting its concentration to suit the current circumstances. 2. Staining suspension cells with cells 2.1 Suspension cells: Centrifuge the cells to collect them, then add PBS and wash twice for five minutes apiece. At 1×106/mL, the cell density 2.2, after staining, add 1 mL of working solution and wait for 5 to 30 minutes. 2.3 Centrifuge for 3–4 minutes at 400 g, then remove the supernatant. 2.4 After adding PBS, wash the cells twice for five minutes each. 2.5 Re-suspend the cells in 1 milliliter of PBS or plasma-free solution, and use a flow cytometer or fluorescence microscope to monitor. 3. Staining adherent cells in cells 3.1 Apply stain to the adhering cells. 3.2 Aspirate extra cells and remove the coverslip from the culture medium. 3.3 Cover the cells completely with 100μL of dye working solution and shake gently for a duration of 5-30 minutes. 3.4 Aspirate the dye working solution, give the cells two to three 5-minute washings, and use a flow cytometer or fluorescence microscope to observe.
RhoNox-1 shows high selectivity for Fe2+ over other divalent metal ions (e.g., Ca2+, Mg2+, Zn2+, Cu2+). In living cells, it exhibits excellent membrane permeability and tends to localize in the Golgi apparatus. Upon reaction with Fe2+, it generates an irreversible orange-red fluorescent product with a large Stokes shift. Ex/Em: 540/575 nm. |
| ln Vivo |
RhoNox-1 is not used for systemic in vivo administration as a therapeutic drug. It is a fluorescent probe for cell imaging studies. The compound can be used for ex vivo imaging of tissue sections to detect Fe2+ levels. It can also be used to explore mechanisms of oxidative stress, inflammation, and diseases linked to dysregulated iron metabolism.
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| Enzyme Assay |
Not applicable. RhoNox-1 is a Fe2+-specific chemical probe that undergoes a chemical reaction, not a biological interaction with a target protein. To determine the reaction kinetics, RhoNox-1 (1-10 uM) is incubated with Fe2+ (0-100 uM) in a buffer (e.g., HEPES, pH 7.4) at 37degC for 0-30 min. Fluorescence intensity (Ex/Em 540/575 nm) is measured.
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| Cell Assay |
Live cells (e.g., HeLa, neuronal cells) are seeded on coverslips or in 96-well plates. Cells are incubated with RhoNox-1 (1-10 uM) in culture medium for 30-60 min at 37degC, then washed. Fe2+ levels are modulated by adding Fe2+ ionophores (e.g., pyrithione) or by transfecting cells with ferritin or DMT1. Fluorescence is visualized by confocal microscopy (Ex/Em 540/575 nm) or measured by flow cytometry.
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| Animal Protocol |
RhoNox-1 is not administered to live animals for in vivo imaging. For ex vivo studies, animals are euthanized, and tissues (e.g., brain, liver) are rapidly dissected, embedded, and sectioned. Tissue sections are incubated with RhoNox-1 (5-20 uM) for 30-60 min, washed, and mounted for fluorescence microscopy to detect Fe2+ levels in specific regions (e.g., substantia nigra in Parkinson‘s disease models).
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| ADME/Pharmacokinetics |
RhoNox-1 is a fluorescent probe, not a drug candidate. Solubility: DMSO (25 mg/mL). Ex/Em maxima: 540/575 nm. Storage: powder at -20degC for 3 years; in solvent at -80degC for 6 months. Protect from light. Molecular formula: C28H30N2O4; molecular weight: 458.55.
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| Toxicity/Toxicokinetics |
Standard safety precautions for chemical probes apply. RhoNox-1 is for research use only. Avoid inhalation and contact. Use PPE in a well-ventilated area. The compound is light-sensitive; protect from light during handling and storage.
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| References |
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| Additional Infomation |
This is a research-use-only fluorescent probe. It is a cell-permeable Fe2+ probe that is also known as FeRhoNox-1. It can be used to study iron metabolism in diseases such as cancer, neurodegenerative disorders, cardiovascular diseases, and to assess therapeutic approaches for diseases linked to excessive ROS production.
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| Molecular Formula |
C28H31N2O4+
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|---|---|
| Molecular Weight |
459.557
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| Exact Mass |
458.22
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| CAS # |
1447815-38-4
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| PubChem CID |
168008618
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| Appearance |
Pink to red solid powder
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| LogP |
4.2
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
34
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| Complexity |
900
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C(C1C=CC=CC=1C1=C2C=CC(N(CC)CC)=CC2=[O+]C2C=C(N(=O)(CC)CC)C=CC1=2)(=O)O
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| InChi Key |
VBBOPWUGWHUQLA-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C28H30N2O4/c1-5-29(6-2)19-13-15-23-25(17-19)34-26-18-20(30(33,7-3)8-4)14-16-24(26)27(23)21-11-9-10-12-22(21)28(31)32/h9-18H,5-8H2,1-4H3
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| Chemical Name |
2-[3-diethylazaniumylidene-6-[diethyl(oxido)azaniumyl]xanthen-9-yl]benzoate
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~218.08 mM)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1760 mL | 10.8800 mL | 21.7599 mL | |
| 5 mM | 0.4352 mL | 2.1760 mL | 4.3520 mL | |
| 10 mM | 0.2176 mL | 1.0880 mL | 2.1760 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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