Z-Arg-Arg-AMC |
Catalog No.GA23711
One-Click Copy Product Info
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Z-Arg-Arg-AMC is a specific substrate for histone B with an Ex/Em of 360/460nm.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 88937-61-5
Sample solution is provided at 25 µL, 10mM.
Z-Arg-Arg-AMC is a specific substrate for histone B with an Ex/Em of 360/460nm. Z-Arg-Arg-AMC has pH-dependent cleavage properties.At neutral pH, Z-Arg-Arg-AMC is readily cleaved by histone B. However, Z-Arg-Arg-AMC is very poorly active at acidic pH with only one-third of the activity at neutral pH [1-4].
References:
[1] Ma C, Liang K, Tang L, He S, Liu X, He M, Li Y. Identification and characteristics of a cathepsin L-like cysteine protease from Clonorchis sinensis. Parasitol Res. 2019 Mar;118(3):829-835.
[2] Yoon MC, Phan V, Podvin S, Mosier C, O'Donoghue AJ, Hook V. Distinct Cleavage Properties of Cathepsin B Compared to Cysteine Cathepsins Enable the Design and Validation of a Specific Substrate for Cathepsin B over a Broad pH Range. Biochemistry. 2023 Aug 1;62(15):2289-2300.
[3] Yoon M C, Hook V, O’Donoghue A J. Cathepsin B dipeptidyl carboxypeptidase and endopeptidase activities demonstrated across a broad pH range[J]. Biochemistry, 2022, 61(17): 1904-1914.
[4] Klemenčič M, Novinec M, Dolinar M. Orthocaspases are proteolytically active prokaryotic caspase homologues: the case of M icrocystis aeruginosa[J]. Molecular Microbiology, 2015, 98(1): 142-150.
This scheme is from the literature, please modify it according to your specific needs.
1. Solution configuration:
(1) Use anhydrous DMSO to dissolve Z-Arg-Arg-AMC to 100mM, and store in small portions at -20℃ to avoid repeated freeze-thaw.
(2) Use PAB (Hank's equilibrium salt solution does not contain sodium bicarbonate, contains 0.6mM CaCl2, 0.6mM MgCl2, 2 mm L-cysteine, 25 mm Pipes) to dilute Z-Arg-Arg-AMC with a concentration of 5-100μM.
Note: Please adjust the concentration of the working liquid according to the actual situation.
2. Cellular assay for cathepsin B activity
(1) Spread cells in 96-well plates and grow to 80% confluence (10-20,000 cells per well).
(2) Media was aspirated and cells were washed with 0.1mL Dulbecco's NaCl/Pi.
(3) Incubate cells with 0.2mL PAB for 30 minutes at 37°C.
(4) The buffer was replaced with fresh PAB containing 100µM Z-Arg-Arg-AMC, and cellular tissue protease B activity was determined in the presence or absence of 0.1% Triton X-100 (which increases the hydrolytic activity of Z-Arg-Arg-AMC).
(5) Detection was performed using a Labsystems Fluoroskan II microplate reader in kinetic mode at 37°C using a 360nm excitation filter and a 460nm emission filter.
Reference:
[1] Hulkower KI, Butler CC, Linebaugh BE, Klaus JL, Keppler D, Giranda VL, Sloane BF. Fluorescent microplate assay for cancer cell-associated cathepsin B. Eur J Biochem. 2000 Jul;267(13):4165-70.
| Cas No. | 88937-61-5 | SDF | |
| Formula | C30H39N9O6 | M.Wt | 621.7 |
| 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 | 1.6085 mL | 8.0425 mL | 16.0849 mL |
| 5 mM | 321.7 μL | 1.6085 mL | 3.217 mL |
| 10 mM | 160.8 μL | 804.2 μL | 1.6085 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 33 reference(s) in Google Scholar.)