| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
Trypsin-like serine proteases are the primary molecular target. The substrate is designed to be specifically cleaved by these enzymes, releasing a fluorescent product.
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| ln Vitro |
Guidelines (This is our suggested protocol; it should be adjusted based on your unique requirements as it just offers as a guideline). Assay for trypsin activity [2] 1. Add 800 μL of 50 mM Tris-HCl buffer (pH 8.0) to 100 μL of diluted trypsin enzyme. 2. Add 100 μL of Boc-Val-Pro-Arg-MCA, a 50 μM substrate, to start the reaction right away. 3. Incubate for ten minutes at 55°C. 4. To halt the reaction, add 1.5 mL of the stop agent (methanol: n-butanol: distilled water = 35:30:35, v/v/v). 5. To evaluate the intensity of fluorescence, use a fluorescence spectrometer (emission 450 nm, excitation 380 nm).
Boc-Val-Pro-Arg-MCA HCl is a sensitive fluorescent substrate for the determination of trypsin-like serine proteases activity. The interaction of Boc-Val-Pro-Arg-MCA with trypsin-like serine proteases results in the production of a fluorescent signal (Ex/Em: 380/450 nm). |
| ln Vivo |
There is no specific in vivo activity data available for this compound. This substrate is typically used in vitro for enzyme activity assays and is not intended for in vivo imaging applications due to potential non-specific cleavage.
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| Enzyme Assay |
A standard protocol involves incubating Boc-Val-Pro-Arg-MCA hydrochloride (e.g., 10-100 uM) with varying concentrations of a trypsin-like serine protease (e.g., trypsin, plasmin) in an assay buffer (e.g., Tris-HCl, pH 8.0) at 37degC. The fluorescence intensity (Ex/Em: 380/450 nm) is measured continuously or after a fixed time.
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| Cell Assay |
A general protocol for cellular experiments involves lysing cells and then incubating the lysate with Boc-Val-Pro-Arg-MCA hydrochloride (e.g., 50 uM) in an assay buffer. The increase in fluorescence over time is measured to determine the trypsin-like protease activity in the sample. A specific inhibitor can be used to confirm specificity.
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| Animal Protocol |
There is no standard in vivo protocol for this substrate. Due to the ubiquity of trypsin-like proteases, in vivo use would likely result in rapid, non-specific cleavage and high background. This product is designed for in vitro biochemical and cell-based assays.
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| ADME/Pharmacokinetics |
General PK properties for this peptide-based substrate are not applicable, as it is not used as a drug or in vivo imaging agent. Formulation is typically in DMSO or assay buffer.
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| Toxicity/Toxicokinetics |
General toxicity for peptide substrates is considered negligible. At working concentrations (uM to mM range), Boc-Val-Pro-Arg-MCA hydrochloride exhibits no significant cytotoxicity. Standard safety precautions for handling laboratory chemicals should be observed.
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| References |
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| Additional Infomation |
Boc-Val-Pro-Arg-MCA is a fluorogenic substrate where the MCA (4-methylcoumaryl-7-amide) group is attached to the arginine residue. Upon cleavage by trypsin-like proteases, free MCA is released, which emits strong fluorescence. This product is for research use only.
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| Molecular Formula |
C31H46CLN7O7
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|---|---|
| Molecular Weight |
664.19
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| Exact Mass |
663.315
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| CAS # |
70375-24-5
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| Related CAS # |
Boc-Val-Pro-Arg-AMC;65147-04-8
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| PubChem CID |
88812723
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| Appearance |
White to off-white solid powder
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| LogP |
5.113
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| Hydrogen Bond Donor Count |
6
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
13
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| Heavy Atom Count |
46
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| Complexity |
1180
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| Defined Atom Stereocenter Count |
3
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| SMILES |
CC1=CC(=O)OC2=C1C=CC(=C2)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H]3CCCN3C(=O)[C@H](C(C)C)NC(=O)OC(C)(C)C.Cl
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| InChi Key |
ROQGJTZGLVLVIN-AAJWHBHYSA-N
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| InChi Code |
InChI=1S/C31H45N7O7.ClH/c1-17(2)25(37-30(43)45-31(4,5)6)28(42)38-14-8-10-22(38)27(41)36-21(9-7-13-34-29(32)33)26(40)35-19-11-12-20-18(3)15-24(39)44-23(20)16-19;/h11-12,15-17,21-22,25H,7-10,13-14H2,1-6H3,(H,35,40)(H,36,41)(H,37,43)(H4,32,33,34);1H/t21-,22-,25-;/m0./s1
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| Chemical Name |
tert-butyl N-[(2S)-1-[(2S)-2-[[(2S)-5-(diaminomethylideneamino)-1-[(4-methyl-2-oxochromen-7-yl)amino]-1-oxopentan-2-yl]carbamoyl]pyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl]carbamate;hydrochloride
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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, avoid exposure to moisture. |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 1.5056 mL | 7.5280 mL | 15.0559 mL | |
| 5 mM | 0.3011 mL | 1.5056 mL | 3.0112 mL | |
| 10 mM | 0.1506 mL | 0.7528 mL | 1.5056 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
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