| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
The primary target of pGlu-Pro-Arg-MNA is serine proteases that recognize the pGlu-Pro-Arg sequence. This sequence is a known substrate motif for thrombin and other coagulation factors. The substrate is cleaved after the arginine residue, releasing free MNA. The rate of MNA release is proportional to protease activity and can be quantified by absorbance at 340 nm (after diazotization) or by fluorescence (excitation 340 nm, emission 425 nm).
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|---|---|
| ln Vitro |
Using reagents such as a chromogenic substrate (pGlu-Pro-Arg-mNA) and a partial thromboplastin time (aPTT) system, protein C activity is determined by photometric analysis [1].
In vitro, pGlu-Pro-Arg-MNA is used to measure the activity of purified proteases or proteases in biological samples. The substrate is incubated with the protease in assay buffer, and MNA release is monitored continuously by absorbance or fluorescence. The assay is used to determine kinetic parameters (Km, Vmax) and to screen for protease inhibitors. It is particularly useful for measuring thrombin activity in plasma or purified systems. |
| ln Vivo |
In vivo activity data for pGlu-Pro-Arg-MNA are not applicable, as it is an in vitro diagnostic substrate. It is not administered to animals. The substrate is used ex vivo to measure protease activity in plasma, tissue homogenates, or other biological samples collected from animals or humans. It serves as an analytical tool for studying protease function in physiological and pathological conditions.
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| Enzyme Assay |
For non-cellular enzyme assays, pGlu-Pro-Arg-MNA is dissolved in DMSO or water and added to protease samples in assay buffer (e.g., Tris-HCl, pH 8.0, containing NaCl and CaCl2). The reaction is monitored continuously at 37°C by measuring absorbance at 340 nm (after diazotization with fast blue B) or fluorescence (excitation 340 nm, emission 425 nm). The initial rate of MNA release is used to calculate enzyme activity.
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| Cell Assay |
For cell-based assays, pGlu-Pro-Arg-MNA is not used directly on cells, as it is a substrate for extracellular or secreted proteases. Cell culture supernatants or cell lysates are collected and incubated with the substrate to measure protease activity. This approach is used to study protease secretion by cells or protease activation in cell lysates. Cell viability is assessed separately by MTT assay.
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| Animal Protocol |
For in vivo experiments, pGlu-Pro-Arg-MNA is not administered to animals. It is used ex vivo to measure protease activity in biological samples. For example, plasma is collected from animals treated with anticoagulants or protease inhibitors, and thrombin activity is measured using the substrate. Tissue homogenates are prepared and assayed for protease activity. The substrate provides a sensitive and specific method for quantifying protease activity in complex biological matrices.
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| References |
[1]. Girolami A, et al. Heterozygous protein-S deficiency: a study of a large kindred. Acta Haematol. 1990;84(3):162-8.
|
| Molecular Formula |
C23H32N8O7
|
|---|---|
| Molecular Weight |
532.55
|
| Exact Mass |
532.239
|
| CAS # |
130835-45-9
|
| Related CAS # |
pGlu-Pro-Arg-MNA monoacetate;2070009-26-4
|
| PubChem CID |
92044478
|
| Sequence |
{pGlu}-Pro-Arg-MNA{pGlu}-PR-MNA
|
| SequenceShortening |
{pGlu}-PR-MNA
|
| Appearance |
Typically exists as solid at room temperature
|
| tPSA |
227
|
| Hydrogen Bond Donor Count |
5
|
| Rotatable Bond Count |
10
|
| Heavy Atom Count |
38
|
| Complexity |
935
|
| Defined Atom Stereocenter Count |
3
|
| SMILES |
COC1=C(C=CC(=C1)[N+](=O)[O-])NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H]2CCCN2C(=O)[C@@H]3CCC(=O)N3
|
| InChi Key |
PWEMRMLJCWKESY-ULQDDVLXSA-N
|
| InChi Code |
InChI=1S/C23H32N8O7/c1-38-18-12-13(31(36)37)6-7-14(18)28-20(33)15(4-2-10-26-23(24)25)29-21(34)17-5-3-11-30(17)22(35)16-8-9-19(32)27-16/h6-7,12,15-17H,2-5,8-11H2,1H3,(H,27,32)(H,28,33)(H,29,34)(H4,24,25,26)/t15-,16-,17-/m0/s1
|
| Chemical Name |
(2S)-N-[(2S)-5-(diaminomethylideneamino)-1-(2-methoxy-4-nitroanilino)-1-oxopentan-2-yl]-1-[(2S)-5-oxopyrrolidine-2-carbonyl]pyrrolidine-2-carboxamide
|
| 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 |
| 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) |
H2O
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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 | 1.8778 mL | 9.3888 mL | 18.7776 mL | |
| 5 mM | 0.3756 mL | 1.8778 mL | 3.7555 mL | |
| 10 mM | 0.1878 mL | 0.9389 mL | 1.8778 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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