| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg | |||
| Other Sizes |
| Targets |
Mca-(Ala7,Lys(Dnp)9)-Bradykinin targets kinin-degrading enzymes, specifically angiotensin-converting enzyme (ACE) and other peptidases that cleave bradykinin. ACE is a key enzyme in the renin-angiotensin system that degrades bradykinin and converts angiotensin I to angiotensin II. This substrate is used to measure ACE activity in vitro by monitoring the increase in fluorescence upon enzymatic cleavage of the peptide bond between the fluorophore and quencher.
|
|---|---|
| ln Vitro |
Enkephalinase also uses Mca-(ala7,lys(dnp)9)-bradykinin as a substrate, but ECE-1 can hydrolyze it ten times more efficiently, making this substrate exclusive to ECE-1 [1].
In vitro, Mca-(Ala7,Lys(Dnp)9)-Bradykinin is used as a fluorogenic substrate to measure kininase and ACE activity. The intact peptide exhibits low fluorescence due to intramolecular quenching between the Mca fluorophore and the Dnp quencher. Upon cleavage by ACE or other peptidases, the quencher is separated from the fluorophore, resulting in increased fluorescence. This assay is used to screen for ACE inhibitors and to study enzyme kinetics in biochemical and pharmacological research. |
| ln Vivo |
In vivo, Mca-(Ala7,Lys(Dnp)9)-Bradykinin is used to study kininase activity and ACE function. However, as a fluorescent substrate, it is primarily used in in vitro assays rather than in vivo. The substrate can be used to measure ACE activity in biological samples such as plasma or tissue homogenates. It is a valuable tool for studying the renin-angiotensin system and for screening potential ACE inhibitors for therapeutic applications.
|
| Enzyme Assay |
In vitro enzyme assays for Mca-(Ala7,Lys(Dnp)9)-Bradykinin involve incubating the fluorogenic substrate with purified ACE or other kininases in appropriate buffer systems. The increase in fluorescence (excitation ~320 nm, emission ~400 nm) is monitored continuously over time using a fluorometer or microplate reader. Enzyme activity is calculated from the initial rate of fluorescence increase. Inhibitors are evaluated by measuring the reduction in activity. Michaelis-Menten kinetics (Km, Vmax) can be determined from substrate concentration curves.
|
| Cell Assay |
In vitro cell-based assays using Mca-(Ala7,Lys(Dnp)9)-Bradykinin are less common as this is primarily a biochemical substrate for enzyme activity measurements. However, the substrate can be used to measure ACE activity in cell culture supernatants or cell lysates. Cells expressing ACE (e.g., endothelial cells) are cultured and the substrate is added to the medium. Fluorescence increase is measured to assess ACE activity secreted or present on the cell surface. ACE inhibitors can be tested for their ability to block cellular ACE activity.
|
| Animal Protocol |
In vivo animal studies using Mca-(Ala7,Lys(Dnp)9)-Bradykinin are limited as it is a fluorescent substrate primarily for in vitro use. The substrate could potentially be used to measure ACE activity in plasma or tissue samples from animal models. For example, plasma from treated animals could be incubated with the substrate to assess ACE activity as a pharmacodynamic marker. However, the substrate itself is not typically administered to animals for in vivo imaging.
|
| ADME/Pharmacokinetics |
Pharmacokinetic properties of Mca-(Ala7,Lys(Dnp)9)-Bradykinin are not relevant for this substrate, as it is used as a reagent in in vitro enzyme assays rather than as a therapeutic or imaging agent. The substrate is stable under recommended storage conditions and is typically dissolved in appropriate buffers for enzyme assays. Stability in biological samples should be considered when measuring ACE activity in plasma or tissue homogenates.
|
| Toxicity/Toxicokinetics |
Toxicity data for Mca-(Ala7,Lys(Dnp)9)-Bradykinin are limited as it is a research reagent used in biochemical assays. The compound is not intended for human therapeutic use. Standard laboratory safety precautions for handling fluorescent compounds and peptides apply. The Dnp group is a potential sensitizer; appropriate personal protective equipment should be used. The substrate is for research use only.
|
| References | |
| Additional Infomation |
Mca-(Ala7,Lys(Dnp)9)-Bradykinin is a fluorogenic peptide substrate for measuring kininase and ACE activity. It contains a Mca fluorophore and a Dnp quencher; cleavage by ACE results in increased fluorescence. This substrate is widely used for screening ACE inhibitors and studying enzyme kinetics in the renin-angiotensin system. It is for research use only and not for human therapeutic or diagnostic use.
|
| Molecular Formula |
C66H81N15O19
|
|---|---|
| Molecular Weight |
1388.43844
|
| Exact Mass |
1387.58
|
| CAS # |
323577-36-2
|
| PubChem CID |
137241787
|
| Appearance |
Light yellow to yellow solid powder
|
| LogP |
5.217
|
| Hydrogen Bond Donor Count |
12
|
| Hydrogen Bond Acceptor Count |
21
|
| Rotatable Bond Count |
34
|
| Heavy Atom Count |
100
|
| Complexity |
2950
|
| Defined Atom Stereocenter Count |
8
|
| SMILES |
C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCCNC2=C(C=C(C=C2)[N+](=O)[O-])[N+](=O)[O-])C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC3=CC=CC=C3)NC(=O)CNC(=O)[C@@H]4CCCN4C(=O)[C@@H]5CCCN5C(=O)[C@H](CCCN=C(N)N)NC(=O)CC6=CC(=O)OC7=C6C=CC(=C7)OC
|
| InChi Key |
FDQLQIBWKJMUSI-HGGPOPJMSA-N
|
| InChi Code |
InChI=1S/C66H81N15O19/c1-38(58(86)76-49(31-40-16-7-4-8-17-40)60(88)75-47(65(93)94)18-9-10-26-69-45-25-22-42(80(95)96)34-53(45)81(97)98)72-61(89)50(37-82)77-59(87)48(30-39-14-5-3-6-15-39)74-56(84)36-71-62(90)51-20-12-28-78(51)64(92)52-21-13-29-79(52)63(91)46(19-11-27-70-66(67)68)73-55(83)32-41-33-57(85)100-54-35-43(99-2)23-24-44(41)54/h3-8,14-17,22-25,33-35,38,46-52,69,82H,9-13,18-21,26-32,36-37H2,1-2H3,(H,71,90)(H,72,89)(H,73,83)(H,74,84)(H,75,88)(H,76,86)(H,77,87)(H,93,94)(H4,67,68,70)/t38-,46-,47-,48-,49-,50-,51-,52-/m0/s1
|
| Chemical Name |
(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-1-[(2S)-1-[(2S)-5-(diaminomethylideneamino)-2-[[2-(7-methoxy-2-oxochromen-4-yl)acetyl]amino]pentanoyl]pyrrolidine-2-carbonyl]pyrrolidine-2-carbonyl]amino]acetyl]amino]-3-phenylpropanoyl]amino]-3-hydroxypropanoyl]amino]propanoyl]amino]-3-phenylpropanoyl]amino]-6-(2,4-dinitroanilino)hexanoic acid
|
| 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: 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)
|
| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~72.02 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 | 0.7202 mL | 3.6012 mL | 7.2023 mL | |
| 5 mM | 0.1440 mL | 0.7202 mL | 1.4405 mL | |
| 10 mM | 0.0720 mL | 0.3601 mL | 0.7202 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
