| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| Other Sizes |
| Targets |
CYP1
CYP1A1, CYP1A2, CYP1B1, NO synthase |
|---|---|
| ln Vitro |
7-Ethoxyresorufin is a fluorometric substrate and competitive inhibitor of cytochrome P450, particularly CYP1A1. It is used as a specific substrate for CYP1A enzymes to measure ethoxyresorufin-O-deethylase (EROD) activity. CYP1A enzymes catalyze the O-deethylation of 7-ethoxyresorufin to resorufin, which is highly fluorescent (excitation ~530 nm, emission ~580 nm). 7-Ethoxyresorufin also inhibits NO synthase. In vitro, 7-Ethoxyresorufin (2-10 uM) binds to the heme moiety of CYP enzymes. It is suitable for studying EROD activity in various biological samples.
|
| ln Vivo |
No specific in vivo data for 7-Ethoxyresorufin as a therapeutic agent; it is a research substrate used to measure CYP1A activity in vivo. In zebrafish, in vitro CYP1A activity has been assessed using 7-ethoxyresorufin as a substrate, showing temporal patterns during organogenesis and lack of gender differences in adult stage. In vivo studies in rodents involve administration of 7-ethoxyresorufin (typically intraperitoneally or orally) followed by measurement of resorufin formation in liver microsomes ex vivo or by imaging fluorescence in tissues. The compound is used to assess the induction of CYP1A enzymes by environmental toxins (e.g., dioxins, PAHs) or drugs.
|
| Enzyme Assay |
EROD activity assay using liver microsomes or recombinant CYP1A enzymes. Microsomes (25-100 ug protein) or recombinant CYP1A1 (1-10 pmol) are incubated with 7-ethoxyresorufin (0.1-10 uM) in a reaction buffer containing potassium phosphate (pH 7.4-8.0), MgCl2, and an NADPH-regenerating system (glucose-6-phosphate, glucose-6-phosphate dehydrogenase, NADP+). The reaction is incubated at 37degC for 5-30 minutes. The reaction is stopped by adding ice-cold methanol or acetonitrile. The fluorescence of the product resorufin is measured using a fluorescence plate reader or fluorometer (excitation 530-550 nm, emission 580-595 nm). A standard curve is prepared using resorufin. Alternatively, resorufin can be quantified by HPLC with fluorescence detection. Kinetic parameters (Km, Vmax) are determined. For inhibition studies, the test compound is pre-incubated with the enzyme before substrate addition.
|
| Cell Assay |
Cell-based EROD assay using primary hepatocytes, HepG2 cells, or other CYP1A-expressing cell lines. Cells are cultured in appropriate media and treated with CYP1A inducers (e.g., 3-methylcholanthrene, beta-naphthoflavone, TCDD) for 24-48 hours to induce CYP1A expression. After induction, cells are washed and incubated with 7-ethoxyresorufin (1-10 uM) in culture medium for 30-60 minutes at 37degC. The supernatant is collected, and the fluorescence of resorufin is measured (excitation 530 nm, emission 585 nm). For kinetic studies, the reaction can be stopped by adding NaOH to the medium. Alternatively, a plate-based kinetic assay is performed: after adding 7-ethoxyresorufin, the increase in fluorescence is measured in real time at 37degC using a fluorescence plate reader. Total protein content or cell number is used to normalize activity. CYP1A protein levels can be confirmed by Western blot. Cell viability is assessed by MTT assay.
|
| Animal Protocol |
For ex vivo EROD activity measurement in animals, rats or mice are dosed with CYP1A inducers (e.g., beta-naphthoflavone, 20-80 mg/kg i.p. for 2-4 days) to induce hepatic CYP1A enzymes. On the day of the experiment, animals are euthanized, and livers are collected. Liver microsomes are prepared by differential centrifugation. Microsomal EROD activity is measured as described in the in vitro enzyme protocol (field 5), using 7-ethoxyresorufin (1-10 uM). Activity is expressed as pmol resorufin/min/mg microsomal protein. For in vivo EROD activity measurement, 7-ethoxyresorufin (5-20 mg/kg) is administered intraperitoneally (i.p.) to rodents. At various time points (e.g., 0.5-4 hours), blood is collected, and plasma is analyzed for resorufin fluorescence. Alternatively, the liver is quickly excised and frozen, and resorufin is extracted from the tissue and quantified by fluorescence. In zebrafish, whole larvae or dissected tissues are incubated with 7-ethoxyresorufin, and fluorescence is measured.
|
| ADME/Pharmacokinetics |
Metabolism / Metabolites
7-Ethoxyhalogenated ... 7-Ethoxyresorufin itself is not a drug; its pharmacokinetics as a probe substrate are not typically studied. When administered in vivo (i.p., 5-20 mg/kg in rodents), 7-ethoxyresorufin is rapidly metabolized by CYP1A enzymes in the liver to resorufin. The plasma concentration of 7-ethoxyresorufin declines rapidly due to extensive metabolism. Resorufin is further conjugated (glucuronidation, sulfation) and excreted in urine and bile. The half-life of 7-ethoxyresorufin is likely very short (minutes) due to rapid metabolism. For research use, the compound is used as an activity-based probe rather than for pharmacokinetic characterization. |
| Toxicity/Toxicokinetics |
7-Ethoxyresorufin is a research reagent and is not intended for human use. Toxicological data are limited; however, as a substrate of CYP1A enzymes, it is expected to have low acute toxicity. Pyrene-based compounds can cause photosensitivity and skin irritation. In zebrafish studies, EROD activity measured with 7-ethoxyresorufin is used as a biomarker of exposure to CYP1A-inducing pollutants (e.g., PAHs, dioxins, PCBs) that are often toxic and carcinogenic. The compound itself is not highly toxic at the concentrations used in EROD assays (1-10 uM in vitro, 5-20 mg/kg in vivo). Handle with standard laboratory precautions; avoid inhalation, skin contact, and ingestion.
|
| References |
|
| Additional Infomation |
7-Ethoxyhalogen is a phenoxazine.
7-Ethoxyresorufin (CAS: 5725-91-7) is a fluorometric substrate and competitive inhibitor of cytochrome P450, particularly CYP1A1, CYP1A2, and CYP1B1. It is used to measure ethoxyresorufin-O-deethylase (EROD) activity, a marker for CYP1A enzyme activity. The deethylation reaction produces resorufin, a highly fluorescent compound, enabling sensitive detection. It is widely used in toxicology, pharmacology, and environmental monitoring to assess CYP1A induction by xenobiotics (e.g., dioxins, PAHs, polychlorinated biphenyls, drugs). The compound also inhibits NO synthase. Molecular formula: C14H11NO3, molecular weight: 241.24. Solubility: soluble to 1 mM in DMSO with gentle warming. Appearance: orange-red solid. Purity: >95%. Synonyms: Resorufin ethyl ether, 7-ER. Storage: 20degC for 3 years; in solvent: -80degC for 1 year. It is suitable for studying EROD activity in zebrafish, rodents, and human tissue samples. For research use only, not for clinical or diagnostic use. |
| Molecular Formula |
C14H11NO3
|
|---|---|
| Molecular Weight |
241.24
|
| Exact Mass |
241.073
|
| CAS # |
5725-91-7
|
| Related CAS # |
7-Ethoxyresorufin-d5;2749329-28-8
|
| PubChem CID |
3294
|
| Appearance |
Orange to red solid powder
|
| Density |
1.3±0.1 g/cm3
|
| Boiling Point |
389.3±42.0 °C at 760 mmHg
|
| Melting Point |
223-225ºC(lit.)
|
| Flash Point |
185.9±22.3 °C
|
| Vapour Pressure |
0.0±0.9 mmHg at 25°C
|
| Index of Refraction |
1.624
|
| LogP |
2.02
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
18
|
| Complexity |
450
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CCOC1=CC2=C(C=C1)N=C3C=CC(=O)C=C3O2
|
| InChi Key |
CRCWUBLTFGOMDD-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C14H11NO3/c1-2-17-10-4-6-12-14(8-10)18-13-7-9(16)3-5-11(13)15-12/h3-8H,2H2,1H3
|
| Chemical Name |
7-ethoxyphenoxazin-3-one
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMF: 2 mg/mL (8.29 mM)
|
|---|---|
| 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 | 4.1452 mL | 20.7262 mL | 41.4525 mL | |
| 5 mM | 0.8290 mL | 4.1452 mL | 8.2905 mL | |
| 10 mM | 0.4145 mL | 2.0726 mL | 4.1452 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
