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100mg | ||
250mg | ||
500mg | ||
Other Sizes |
ln Vitro |
Assay for CYP3A4 activity[2]: 1. Get the reaction solution ready. a. The fluorescent substrate, Resorufin benzyl ether, has a concentration of 1 mM in the standard stock solution. 500 mL of dimethyl sulfoxide, 3.5 mL of acetonitrile, 1 mL of 2% w/v Poloxamer 188, and 5 mg of resorufin benzyl ether should all be dissolved. b. Make a new solution of the CYP3A4 enzyme. To create a 5 nM enzyme solution, dilute 5 mL of the 1 mM enzyme stock solution with 995 mL of buffer. 2. Find out what CYP3A4 activity is. In a 96-well plate, react. Fill each well with 1 mL of 1 mM Resorufin benzyl ether and 99 mL of buffer mix. Adjust to 5 mM as the final concentration. b. After transferring 100 mL of the 5 nM enzyme solution, incubate for 30 minutes at 37°C. C. Measure the enzyme activity using fluorescence detection at the excitation wavelength (lex=570 nm) and emission wavelength (lem=590 nm). d. variables influencing CYP3A4 activity measurement. phosphate and Tris-HCl buffers, for example; also, buffer concentrations (50–200 mM) and incubation times (0–50 min). Measuring the metabolic activity of CYP3A4 [3]: 1. At a final concentration of 5 pmol/well, add the CYP3A4 enzyme. 2. For every reaction, use 200 mM potassium phosphate buffer and 50 pM substrate. < br /> 3. Measure the metabolite fluorescence after 45 minutes of incubation with BzRes. 590 nm is the emission wavelength (Em) while 530 nm is the excitation wavelength (Ex). 4. Indicate the full strength extract concentration as 100% (diluted 1:4 in the final assay volume). 5. Double-do the 100% extract dilution at a ratio of 1:3. Determine the fluorescence values' standard deviation and mean.
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References |
[1]. Ji X, et al. Regulating the activity of boronate moiety to construct fluorescent probes for the detection of ONOO-in vitro and in vivo. Anal Methods. 2022 Dec 15;14(48):5027-5033.
[2]. Nuchtavorn N, et al. Paper-based sol-gel thin films immobilized cytochrome P450 for enzyme activity measurement. Anal Chim Acta. 2020 Feb 15;1098:86-93. [3]. Yale SH, et al. Analysis of the inhibitory potential of Ginkgo biloba, Echinacea purpurea, and Serenoa repens on the metabolic activity of cytochrome P450 3A4, 2D6, and 2C9. J Altern Complement Med. 2005 Jun;11(3):433-9. |
Molecular Formula |
C19H13NO3
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Molecular Weight |
303.31
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CAS # |
87687-02-3
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Appearance |
Typically exists as solids (or liquids in special cases) at room temperature
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SMILES |
O=C1C=C2C(=NC3C(O2)=CC(OCC2C=CC=CC=2)=CC=3)C=C1
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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 |
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 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.) |
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Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
1 mM | 3.2970 mL | 16.4848 mL | 32.9696 mL | |
5 mM | 0.6594 mL | 3.2970 mL | 6.5939 mL | |
10 mM | 0.3297 mL | 1.6485 mL | 3.2970 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.