Tos-Gly-Pro-Arg-AMC . HCl |
Catalog No.GA23623
One-Click Copy Product Info
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Tos-Gly-Pro-Arg-AMC . HCl is a fluorescent substrate for thrombin, which is widely used to detect serine protease activity. Its excitation and emission wavelengths are 380nm and 460nm, respectively.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 117961-27-0
Sample solution is provided at 25 µL, 10mM.
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Tos-Gly-Pro-Arg-AMC . HCl is a fluorescent substrate for thrombin, which is widely used to detect serine protease activity. Its excitation and emission wavelengths are 380nm and 460nm, respectively.
The core structure of Tos-Gly-Pro-Arg-AMC . HCl contains a fluorescent group 7-amino-4-methylcoumarin (AMC). When the substrate is hydrolyzed by protease, AMC is released, generating a strong fluorescent signal (excitation wavelength 380nm, emission wavelength 460nm). The Gly-Pro-Arg sequence is a specific recognition site for thrombin, so this substrate is highly selective for thrombin and is also suitable for the study of other serine proteases (such as trypsin, kallikrein, etc.). Tos-Gly-Pro-Arg-AMC . HCl can also be used for high-throughput screening of thrombin inhibitors or for detecting thrombin activity in blood samples to assist in the diagnosis of coagulation abnormalities[1][2].
References:
[1]. Corvo I, Cancela M, Cappetta M, et al. The major cathepsin L secreted by the invasive juvenile Fasciola hepatica prefers proline in the S2 subsite and can cleave collagen[J]. Molecular and biochemical parasitology, 2009, 167(1): 41-47.
[2]. Wirsching F, Luge C, Schwienhorst A. Modular design of a novel chimeric protein with combined thrombin inhibitory activity and plasminogen-activating potential[J]. Molecular genetics and metabolism, 2002, 75(3): 250-259.
This protocol is only for experimental guidance. The actual application must be modified according to your actual needs.
Trypsin-like activity test in colon supernatant[1][2].
1. Solution preparation
Substrate solution: Dissolve Tos-Gly-Pro-Arg-AMC . HCl in 50mM Tris-HCl (pH 8.0) buffer to prepare a 10mM stock solution. Dilute to a working concentration of 100µM before use and preheat to room temperature.
2. Sample preparation
A: Take 50mg of distal colon sample, rinse with Krebs solution, freeze quickly and store at -80°C.
B: Use liquid nitrogen to pulverize the sample to maintain enzyme activity, add lysis buffer to dissolve for 15min, centrifuge and collect the supernatant for use.
3. Fluorescence detection
A: Place the colon supernatant in a cold microtiter plate, ensuring that the sample remains cold to avoid loss of enzyme activity.
B: Add substrate solution preheated to room temperature and incubate for 20min.
C: Use an ELISA reader to dynamically detect the fluorescence intensity changes of AMC under the conditions of excitation wavelength 380nm and emission wavelength 460nm, and calculate the enzyme activity.
Notes:
1) For your safety and health, please wear lab coats and protective gloves when operating.
2) Enzyme activity experiments are affected by factors such as ion concentration, reaction temperature and pH. It is recommended to explore the optimal concentration when using for the first time.
References:
[1]. Ceuleers H, Hanning N, Heirbaut J, et al. Newly developed serine protease inhibitors decrease visceral hypersensitivity in a post?inflammatory rat model for irritable bowel syndrome[J]. British Journal of Pharmacology, 2018, 175(17): 3516-3533.
[2]. Corvo I, Cancela M, Cappetta M, et al. The major cathepsin L secreted by the invasive juvenile Fasciola hepatica prefers proline in the S2 subsite and can cleave collagen[J]. Molecular and biochemical parasitology, 2009, 167(1): 41-47.
| Cas No. | 117961-27-0 | SDF | |
| Formula | C30H37N7O7S · HCl | M.Wt | 676.19 |
| Solubility | Soluble in DMSO | 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 | 1.4789 mL | 7.3944 mL | 14.7887 mL |
| 5 mM | 295.8 μL | 1.4789 mL | 2.9577 mL |
| 10 mM | 147.9 μL | 739.4 μL | 1.4789 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: >98.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 15 reference(s) in Google Scholar.)