Z-Ala-Ala-Asn-AMC (Synonyms: Cbz-Ala-Ala-Asn-AMC) |
Catalog No.GA23674
One-Click Copy Product Info
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Z-Ala-Ala-Asn-AMC (Cbz-Ala-Ala-Asn-AMC) is a synthetic fluorogenic peptide substrate that releases highly fluorescent AMC upon cleavage by proteases (Ex/Em: 380nm/430nm), which is widely used for the detection of asparaginyl endopeptidase (Legumain) activity.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 149697-16-5
Sample solution is provided at 25 µL, 10mM.
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Z-Ala-Ala-Asn-AMC (Cbz-Ala-Ala-Asn-AMC) is a synthetic fluorogenic peptide substrate that releases highly fluorescent AMC upon cleavage by proteases (Ex/Em: 380nm/430nm), which is widely used for the detection of asparaginyl endopeptidase (Legumain) activity[1,2,3]. Legumain is often overexpressed in tumors[4], and Z-Ala-Ala-Asn-AMC serves as a substrate suitable for detecting and quantifying legumain activity in biological samples, high-throughput inhibitor screening, and enzyme kinetics studies[5,6,7].
References:
[1] ZHAO T, LI Z, GUO Z, et al. Functional recombinant human Legumain protein expression in Pichia pastoris to enable screening for Legumain small molecule inhibitors[J]. Protein Expression and Purification, 2018, 150: 12-16.
[2] POREBA M. Recent advances in the development of legumain-selective chemical probes and peptide prodrugs[J]. Biological Chemistry, 2019, 400(12): 1529-1550.
[3] PICKERING A M, DAVIES K J A. A simple fluorescence labeling method for studies of protein oxidation, protein modification, and proteolysis[J]. Free Radical Biology and Medicine, 2012, 52(2): 239-246.
[4] LIU C, SUN C, HUANG H, et al. Overexpression of legumain in tumors is significant for invasion/metastasis and a candidate enzymatic target for prodrug therapy[J]. Cancer Research, 2003, 63(11): 2957-2964.
[5] ZHAO T, LIU Y, HAO Y, et al. Esomeprazole inhibits the lysosomal cysteine protease legumain to prevent cancer metastasis[J]. Investigational New Drugs, 2021, 39(2): 337-347.
[6] POREBA M, SOLBERG R, RUT W, et al. Counter selection substrate library strategy for developing specific protease substrates and probes[J]. Cell Chemical Biology, 2016, 23(8): 1023-1035.
[7] GRUNCLOVA L, HORN M, VANCOVÁ M, et al. Two secreted cystatins of the soft tick Ornithodoros moubata: differential expression pattern and inhibitory specificity[J]. Biological Chemistry, 2006, 387(12).
Using Z-Ala-Ala-Asn-AMC to determine recombinant Legumain activity[1]:
(1) 10μL aliquots of Legumain (0.6mg/mL) were mixed with 50μL of different activation buffers (containing 50mM Sodium Acetate and 100mM NaCl, pH 4) and pre-incubated at 37℃ for 2h respectively.
(2) Samples were then diluted to 1ng/μL in assay buffer (50mM MES, 250mM NaCl, pH 5).
(3) 30μL of 150μM Z-Ala-Ala-Asn-AMC was added and the enzymatic activity measured using a microplate reader at 380 and 430nm.
(4) Readings were taken every 2min for 20 cycles respectively. One unit of enzyme activity was defined as the activity required to release 1μM of AMC per minute at 37℃.
References:
[1] ZHAO T, LI Z, GUO Z, et al. Functional recombinant human Legumain protein expression in Pichia pastoris to enable screening for Legumain small molecule inhibitors[J]. Protein Expression and Purification, 2018, 150: 12-16.
| Cas No. | 149697-16-5 | SDF | |
| Synonyms | Cbz-Ala-Ala-Asn-AMC | ||
| Formula | C28H31N5O8 | M.Wt | 565.58 |
| 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 | 1.7681 mL | 8.8405 mL | 17.681 mL |
| 5 mM | 353.6 μL | 1.7681 mL | 3.5362 mL |
| 10 mM | 176.8 μL | 884 μL | 1.7681 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 10 reference(s) in Google Scholar.)