Mca-Ala-Pro-Lys(Dnp)-OH |
Katalog-Nr.GA23174
One-Click Copy Product Info
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Mca-Ala-Pro-Lys(Dnp)-OH, ein spezifisches ACE2-gequenchtes fluorogenes Substrat, kann verwendet werden, um ACE2-AktivitÄt wie Urin, Herz und Lunge nachzuweisen.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 305336-82-7
Sample solution is provided at 25 µL, 10mM.
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Mca-Ala-Pro-Lys(Dnp)-OH is a specific quenched fluorogenic substrate for angiotensin-converting enzyme 2 (ACE2). Mca-Ala-Pro-Lys(Dnp)-OH can be used to detect ACE2 activity in biological samples such as urine, heart, and lung tissues. Mca-Ala-Pro-Lys(Dnp)-OH serves as a critical tool in research related to the renin-angiotensin system (RAS), cardiovascular diseases, and pulmonary conditions[1].
References:
[1] Xiao F, Burns KD. Measurement of Angiotensin Converting Enzyme 2 Activity in Biological Fluid (ACE2). Methods Mol Biol. 2017;1527:101-115.
ACE2 Activity Assay in Biological Fluids Using Mca-Ala-Pro-Lys(Dnp)-OH
This protocol is adapted from research data and provided for reference only. Adjustments may be necessary based on specific experimental requirements.
1.Reagent Preparation
(1) ACE2 Substrate Solution: Dilute Mca-Ala-Pro-Lys(Dnp)-OH (15mM stock solution in 1% NH₄OH) with protease inhibitors NEM (100mM) and PMSF (100mM) in pre-chilled MES buffer (50mM MES, 300mM NaCl, 10μM ZnCl₂, pH=6.8) to final concentrations of 15μM, 1mM, and 1mM, respectively.
(2) ACE2 Standards: Reconstitute recombinant human or mouse ACE2 in PBS to 10μg/mL, followed by serial dilution to generate a standard curve ranging from 1.56 to 50ng/mL.
(3) ACE2 Inhibitors: Prepare MLN-4760 (10μM) or DX600 (10μM) for specificity validation.
2.Biological Sample Pretreatment
(1) Urine: Collect samples on ice, centrifuge (12,000×g, 5min, 4°C), aliquot supernatant, and store at -80°C. Dilute samples at least 1:2 with assay buffer before use.
(2) Plasma: Separate plasma from heparin-anticoagulated blood by centrifugation (3,000×g, 10min, 4°C), aliquot, and store at -80°C. Dilute 1:7.5 prior to assay.
(3) Cell Culture Supernatant: Centrifuge serum-free conditioned medium and use the supernatant directly without dilution.
3.ACE2 Activity Detection Procedure
(1) Add 10μL of inhibitor (MLN-4760 or DX600) or ddH₂O (control) to each well of a black 96-well plate.
(2) Add 75μL of ACE2 substrate/buffer mixture to all wells. Add 15μL of diluted biological sample or ACE2 standard to each well, achieving a total reaction volume of 100μL.
(3) Seal the plate with adhesive film and aluminum foil to protect from light. Incubate with shaking at room temperature for 16 hours.
(4) Measure fluorescence using a microplate reader (excitation: 320nm, emission: 405nm) and record relative fluorescence units (RFU).
Precautions:
(1) Samples with high background fluorescence (e.g., concentrated urine) require further dilution. Glucose, urea, creatinine, and albumin do not interfere with assay results.
(2) Avoid using EDTA-containing blood collection tubes or serum (which contains endogenous ACE2 activity). Maintain pH at 6.8±0.2 throughout the assay.
(3) For your safety and health, please wear a lab coat and disposable gloves during operation.
References:
[1] Xiao F, Burns KD. Measurement of Angiotensin Converting Enzyme 2 Activity in Biological Fluid (ACE2). Methods Mol Biol. 2017;1527:101-115.
| Cas No. | 305336-82-7 | SDF | |
| Formula | C32H36N6O12 | M.Wt | 696.67 |
| Löslichkeit | Soluble in water at 25°C, 1,00 g/L | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.4354 mL | 7.177 mL | 14.354 mL |
| 5 mM | 287.1 μL | 1.4354 mL | 2.8708 mL |
| 10 mM | 143.5 μL | 717.7 μL | 1.4354 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.50% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 9 reference(s) in Google Scholar.)