Boc-LRR-AMC (Synonyms: Boc-Leu-Arg-Arg-AMC, Boc-Leu-Arg-Arg-7-amido-4-Methylcoumarin) |
| カタログ番号GC42958 |
Boc-LRR-AMCは、26Sプロテアソームまたは20Sプロテオリチックコアのトリプシン様活性のための蛍光基質です。
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 109358-46-5
Sample solution is provided at 25 µL, 10mM.
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Boc-LRR-AMC is a fluorescent substrate for Trypsin, similar to the 20S proteasome. Boc-LRR-AMC can be used to detect Trypsin-like activity in the 26S proteasome or 20S proteasome core. The detection wavelength of Boc-LRR-AMC is 380nm/460nm[1].
References:
[1]. Gruba N, Wysocka M, Brzezińska M, et al. Bladder cancer detection using a peptide substrate of the 20S proteasome[J]. The FEBS journal, 2016, 283(15): 2929-2948.
CAPA for capture proteasome assay (from the literature, please modify according to your needs) [1][2].
1. Antibody coating: Prepare 5μg/ml of the target antibody to coat a black 96-well maxisorp plate, which is then further blocked for 1 hour using PBS containing 2% BSA.
2. Proteasome capture: Cell pellets were washed in PBS and lysed on ice at a cell density of 107 cells/ml in lysis buffer (50mM TRIS, 0.1% NP40, pH 7.5). The supernatant was collected, analyzed using the BCA protein assay reagent, and the protein content was adjusted to a concentration of 200μg/ml in lysis buffer. 50μl of cell lysate was then added to each well, and the plate was incubated at 4°C for 2 hours. The amount of proteasomes captured under these conditions was adjusted to approximately 200ng per well using a quantitative ELISA assay [3].
3. Substrate incubation: After capturing the proteasomes, the plates were first carefully washed with 20mM Tris buffer (containing 0.1% NP40, pH 7.5) and then washed with 20mM Tris buffer (pH 7.5) to remove excess material. Proteasome inhibitors diluted in 20mM Tris buffer (pH 7.5) were then added to the wells and incubated with rotation at room temperature for 10 minutes. Finally, the fluorescent substrate Boc-LRR-AMC was diluted in 20mM Tris buffer and added to the wells at a concentration of 100μM.
4. Fluorescence detection: The plates were sealed with a plastic cover and incubated at 37°C for approximately 30 minutes. Fluorescence values were then measured using a Proteasome-GloMAX or SpectraMax 190 microplate reader at an excitation wavelength of 380nm and an emission wavelength of 460nm.
References:
[1]. Vigneron N, Abi Habib J, Van den Eynde B J. The capture proteasome assay (CAPA) to evaluate subtype-specific proteasome inhibitors[J]. Data in Brief, 2015, 4: 146-151.
[2]. Vigneron N, Abi Habib J, Van den Eynde B J. The capture proteasome assay: a method to measure proteasome activity in vitro[J]. Analytical biochemistry, 2015, 482: 7-15.
[3]. Guillaume B, Chapiro J, Stroobant V, et al. Two abundant proteasome subtypes that uniquely process some antigens presented by HLA class I molecules[J]. Proceedings of the National Academy of Sciences, 2010, 107(43): 18599-18604.
| Cas No. | 109358-46-5 | SDF | |
| 同義語 | Boc-Leu-Arg-Arg-AMC, Boc-Leu-Arg-Arg-7-amido-4-Methylcoumarin | ||
| Canonical SMILES | O=C1C=C(C)C2=C(C=C(NC([C@H](CCCNC(N)=N)NC([C@H](CCCNC(N)=N)NC([C@H](CC(C)C)NC(OC(C)(C)C)=O)=O)=O)=O)C=C2)O1 | ||
| Formula | C33H52N10O7 | M.Wt | 700.8 |
| 溶解度 | DMF: 14 mg/ml,DMSO: 12 mg/ml,Ethanol: 10 mg/ml,PBS (pH 7.2): 1 mg/ml | 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 | 1.4269 mL | 7.1347 mL | 14.2694 mL |
| 5 mM | 0.2854 mL | 1.4269 mL | 2.8539 mL |
| 10 mM | 0.1427 mL | 0.7135 mL | 1.4269 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
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μL
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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 ddH2O, 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%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 32 reference(s) in Google Scholar.)
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