H-Gln-AMC |
Katalog-Nr.GA22334
One-Click Copy Product Info
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H-Gln-AMC is a fluorogenic substrate commenly used to measure the activity of glutaminyl cyclase (QC) enzymes with Km value of 51 ± 3µM at pH 8.0 and kcat value of 5.4 ± 0.1s-1.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 105888-45-7
Sample solution is provided at 25 µL, 10mM.
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H-Gln-AMC is a fluorogenic substrate commenly used to measure the activity of glutaminyl cyclase (QC) enzymes with Km value of 51 ± 3µM at pH 8.0 and kcat value of 5.4 ± 0.1s-1[1]. Glutaminyl cyclase (QC) catalyzes the intramolecular cyclization of N-terminal glutaminyl (Gln) and glutamyl (Glu) residues to form pyroglutamic (pGlu) peptides or proteins. Some recent findings suggest that QC can be a potential target against Alzheimer’s disease, Huntington disease and other serious neurodegenerative disease[2].
H-Gln-AMC was used as a fluorescent substrate to detect glutaminyl cyclase (QC) activity in cerebrospinal fluid (CSF) from multiple sclerosis (MS) patients. In CSF, H-Gln-AMC was catalyzed by QC to form pyroglutamyl-AMC, which was subsequently hydrolyzed by pyroglutamyl aminopeptidase to release the fluorescent product AMC. The results show a slight increase in QC activity observed in MS patients, although the difference was not significant[3]. H-Gln-AMC was used as a fluorescent substrate to measure the activity of glutaminyl cyclases (QCs) from Porphyromonas gingivalis (PgQC), Tannerella forsythia (TfQC), and Prevotella intermedia (PiQC). At a concentration of 50μM H-Gln-AMC, the catalytic activity of PgQC, TfQC, and PiQC could be accurately determined based on changes in AMC fluorescence intensity[4].
References:
[1] Schilling S, Hoffmann T, Rosche F, et al. Heterologous expression and characterization of human glutaminyl cyclase: evidence for a disulfide bond with importance for catalytic activity. Biochemistry. 2002 Sep 3;41(35):10849-57.
[2] Szaszkó M, Hajdú I, Flachner B, et al. Identification of potential glutaminyl cyclase inhibitors from lead-like libraries by in silico and in vitro fragment-based screening. Mol Divers. 2017.21:175–186
[3] Gontsarova A, Kaufmann E, Tumani H, et al. Glutaminyl cyclase activity is a characteristic feature of human cerebrospinal fluid. Clin Chim Acta. 2008 Mar;389(1-2):152-9.
[4] Taudte N, Linnert M, Rahfeld J, et al.Mammalian-like type II glutaminyl cyclases in Porphyromonas gingivalis and other oral pathogenic bacteria as targets for treatment of periodontitis. J Biol Chem. 2021 Jan-Jun:296:100263.
H-Gln-AMC used as the substrate for QC activity assays.
1. Prepare a 50mM Tris-HCl buffer with 150mM NaCl at pH 8.0.
2. Prepare H-Gln-AMC solutions with varying concentrations (e.g 50µM for bacterial QC catalytic activity testing) in the Tris-HCl buffer.
3. Add 0.5U of pyroglutaminyl aminopeptidase to each well of a 96-well microtiter plate containing the H-Gln-AMC solutions.
4. Incubate the plate at 30°C for 10 minutes.
5. Add approximately 50nM of the QC enzyme to a final volume of 125μL reaction mixture to initiate the reaction.
6. Monitor the fluorescence intensity at excitation/emission wavelengths of 380/460nm using the FluoStar Optima
7. Calculate QC activity from a standard curve of the fluorophore AMC generated under the same assay conditions
This protocol only provides a guideline, and should be modified according to your specific needs.
References:
[1]Taudte N, Linnert M, Rahfeld J, et al.Mammalian-like type II glutaminyl cyclases in Porphyromonas gingivalis and other oral pathogenic bacteria as targets for treatment of periodontitis. J Biol Chem. 2021 Jan-Jun:296:100263.
| Cas No. | 105888-45-7 | SDF | |
| Formula | C15H17N3O4 | M.Wt | 303.32 |
| Löslichkeit | 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 | 3.2968 mL | 16.4842 mL | 32.9685 mL |
| 5 mM | 659.4 μL | 3.2968 mL | 6.5937 mL |
| 10 mM | 329.7 μL | 1.6484 mL | 3.2968 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 36 reference(s) in Google Scholar.)