| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
RETF-4NA TFA targets the enzyme chymase, a serine protease. The substrate is designed to be specifically cleaved by chymase at the Phe-4NA bond. Upon enzymatic cleavage, the 4-nitroaniline (4NA) group is released, generating a yellow color. The TFA salt is the standard form for solubility and stability. It is used as a sensitive and selective substrate for chymase when free or bound to alpha2M.
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| ln Vitro |
In vitro, RETF-4NA TFA is a chromogenic peptide that serves as a sensitive and selective substrate for chymase. It is widely used in enzymology, drug discovery, and protease research to study catalytic mechanisms, inhibitor screening, and pathway regulation. The release of 4-nitroaniline upon cleavage produces a chromogenic signal measurable at 405 nm.
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| Enzyme Assay |
A standard chymase activity assay is performed in a 96-well plate format. Recombinant human chymase (0.1-1 microg) is incubated with varying concentrations of RETF-4NA TFA (25-1000 microM) in assay buffer (50 mM Tris-HCl, pH 8.0, 0.1 M NaCl, 0.05% Brij-35, 100 microg/mL BSA) at 37degC for 10-20 min. The absorbance at 405 nm is measured continuously or as an endpoint. The initial rate of absorbance increase is proportional to enzyme activity. Kinetic parameters (Km, Vmax) are calculated by fitting the data to the Michaelis-Menten equation. For inhibitor studies, the compound is pre-incubated with chymase for 5-10 min before substrate addition.
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| Cell Assay |
RETF-4NA TFA is not used in cell-based assays; it is an in vitro substrate. To measure chymase activity in cell lysates or tissue homogenates, the sample (20-50 ug protein) is added to a 96-well plate. RETF-4NA TFA (100-500 uM) is added to initiate the reaction. The plate is incubated at 37degC for 30-60 min. Absorbance at 405 nm is measured. For inhibitor screening, potential inhibitors are pre-incubated with the lysate for 5-10 min. The IC50 is calculated from the concentration-response curve.
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| Animal Protocol |
RETF-4NA TFA is not used in animal studies. However, to measure chymase activity in vivo, animals (e.g., mice) can be injected with a chymase inhibitor or agonist, then tissues (e.g., lung, skin) are collected and homogenized. Chymase activity in the homogenate is measured using the substrate as described above. Alternatively, the substrate can be used to measure chymase activity in bronchoalveolar lavage fluid (BALF) or plasma collected from animals.
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| ADME/Pharmacokinetics |
RETF-4NA TFA has a molecular weight of 827.76 and the molecular formula C34H44F3N9O12. The product has a purity of ≥99%. It should be stored as a powder at -20degC, protected from light and moisture. In solvent, it is stable for 1 year at -80degC. The TFA salt ensures water solubility. Specific PK parameters have not been reported.
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| Toxicity/Toxicokinetics |
RETF-4NA TFA is generally considered non-toxic for routine laboratory use. As a peptide substrate, it is not intended for in vivo administration. The 4-nitroaniline release product is a mild irritant but is produced only in small amounts in enzymatic assays. Standard safety precautions for handling peptide-based reagents should be followed, including the use of gloves and eye protection.
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| References | |
| Additional Infomation |
RETF-4NA TFA is a research-grade chromogenic substrate and is not approved for clinical use. It is a chymase-specific substrate used for enzyme activity assays, inhibitor screening, and protease research. This product is for research use only and not for human therapeutic applications. Store as a powder at -20degC. The TFA salt form enhances solubility.
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| Molecular Formula |
C34H44F3N9O12
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|---|---|
| Molecular Weight |
827.76
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| Related CAS # |
RETF-4NA;1160928-63-1
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| Appearance |
White to off-white solid powder
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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: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 :~110 mg/mL (~132.89 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.75 mg/mL (3.32 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 27.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.75 mg/mL (3.32 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 27.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.75 mg/mL (3.32 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.2081 mL | 6.0404 mL | 12.0808 mL | |
| 5 mM | 0.2416 mL | 1.2081 mL | 2.4162 mL | |
| 10 mM | 0.1208 mL | 0.6040 mL | 1.2081 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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