Z-Gly-Gly-Arg-AMC acetate |
Catalog No.GC37965
One-Click Copy Product Info
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Z-Gly-Gly-Arg-AMC acetate is a specific thrombin fluorescent substrate with λexc = 355nm, λem = 460nm.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 2070009-61-7
Sample solution is provided at 25 µL, 10mM.
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Z-Gly-Gly-Arg-AMC acetate is a specific thrombin fluorescent substrate with λexc = 355nm, λem = 460nm[1]. Z-Gly-Gly-Arg-AMC acetate acts as the reference substrate in the Calibrated Automated Thrombogram (CAT) for measuring endogenous thrombin potential (ETP) in platelet-rich (PRP) or platelet-poor plasma (PPP)[2]. Z-Gly-Gly-Arg-AMC acetate is employed in 96-well format for rapid identification and potency ranking of direct thrombin inhibitors or protease-activated receptor antagonists[3].
References:
[1] Tarandovskiy I D, Artemenko E O, Panteleev M A, et al. Antiplatelet agents can promote two-peaked thrombin generation in platelet rich plasma: mechanism and possible applications[J]. PLoS One, 2013, 8(2): e55688.
[2] Machlus K R, Colby E A, Wu J R, et al. Effects of tissue factor, thrombomodulin and elevated clotting factor levels on thrombin generation in the calibrated automated thrombogram[J]. Thrombosis and haemostasis, 2009, 102(11): 936-944.
[3] Kale S S, Bergeron-Brlek M, Wu Y, et al. Thiol-to-amine cyclization reaction enables screening of large libraries of macrocyclic compounds and the generation of sub-kilodalton ligands[J]. Science advances, 2019, 5(8): eaaw2851.
Detection of thrombin activity based on Z-Gly-Gly-Arg-AMC acetate[1]
This protocol only provides a guideline, and should be modified according to your specific needs.
1.Solution preparation
(1) Dissolve the Z-Gly-Gly-Arg-AMC acetate solid in an appropriate amount of DMSO to prepare a 50mM stock solution. Store stock solution at -20°C and avoid repeated freezing and thawing.
(2) Dilute the appropriate Z-Gly-Gly-Arg-AMC acetate stock solution in PBS to prepare a 2.5mM working solution. The working solution should be prepared immediately and used up promptly.
(3) Take an appropriate CaCl2 solution and dissolve in the detection buffer (containing 20mM HEPES, 145mM NaCl, pH 7.5) to prepare the activation buffer.
2. Thrombin generation assay
(1) Mix the plasma with the Z-Gly-Gly-Arg-AMC acetate stock solution at a volume ratio of 4:1, and place the mixture in the wells of a flat-bottom 96-well plate. Add 100µl of the mixture to each well. Add 20µl of activation buffer to each well to trigger the clotting reaction. The final concentration of Z-Gly-Gly-Arg-AMC acetate is 400µM.
(2) Fluorescence is continuously measured at 37°C using a fluorescence plate reader (Ex/Em=355/460nm) for a period of 30min with a read every 3min.
3. Notes for Attention
(1) Z-Gly-Gly-Arg-AMC acetate is photosensitive and the operation process should be light-protected.
(2) To avoid bubbles, the reading should be taken within 10 seconds after the activation buffer (containing CaCl₂) is added.
References:
[1] Tarandovskiy I D, Artemenko E O, Panteleev M A, et al. Antiplatelet agents can promote two-peaked thrombin generation in platelet rich plasma: mechanism and possible applications[J]. PLoS One, 2013, 8(2): e55688.
| Cas No. | 2070009-61-7 | SDF | |
| Formula | C30H37N7O9 | M.Wt | 639.66 |
| Solubility | Water : ≥ 60 mg/mL (93.80 mM) | 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 | 1.5633 mL | 7.8167 mL | 15.6333 mL |
| 5 mM | 312.7 μL | 1.5633 mL | 3.1267 mL |
| 10 mM | 156.3 μL | 781.7 μL | 1.5633 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 37 reference(s) in Google Scholar.)