Boc-Phe-Ser-Arg-AMC |
Katalog-Nr.GA21114
One-Click Copy Product Info
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Boc-Phe-Ser-Arg-AMC is a fluorescent substrate that, upon cleavage by proteases, releases highly fluorescent AMC (Ex/Em: 355 nm/460 nm). Boc-Phe-Ser-Arg-AMC is characterized by its preferential hydrolysis by coagulation factor XIa and is widely used for detecting the activities of enzymes such as trypsin, tryptase, and kallikrein.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 73554-90-2
Sample solution is provided at 25 µL, 10mM.
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Boc-Phe-Ser-Arg-AMC is a fluorescent substrate that, upon cleavage by proteases, releases highly fluorescent AMC (Ex/Em: 355 nm/460 nm). Boc-Phe-Ser-Arg-AMC is characterized by its preferential hydrolysis by coagulation factor XIa and is widely used for detecting the activities of enzymes such as trypsin, tryptase, and kallikrein[1,2,3]. Boc-Phe-Ser-Arg-AMC is also highly sensitive to proteases in yeast mitochondrial matrix and trypsin in rat mast cells. After the Arg-AMC peptide bond is cleaved by proteases, AMC is released, and the increase in fluorescence intensity is proportional to the enzyme activity[4,5,6].
References:
[1] KALE S S, BERGERON-BRLEK M, WU Y, et al. Thiol-to-amine cyclization reaction enables screening of large libraries of macrocyclic compounds and the generation of sub-kilodalton ligands[J]. Science Advances, 2019, 5(8): eaaw2851.
[2] GONG Z, DAI S, JIANG X, et al. Variants in KLK11, affecting signal peptide cleavage of kallikrein-related peptidase 11, cause an autosomal-dominant cornification disorder[J]. British Journal of Dermatology, 2023, 188(1): 100-111.
[3] KOMATSU N, SAIJOH K, KUK C, et al. Aberrant human tissue kallikrein levels in the stratum corneum and serum of patients with psoriasis: dependence on phenotype, severity and therapy[J]. British Journal of Dermatology, 2007, 156(5): 875-883.
[4] BRAGANZA V J, SIMMONS W H. Tryptase from rat skin: purification and properties[J]. Biochemistry, 1991, 30(20): 4997-5007.
[5] YASUHARA T, OHASHI A. New chelator-sensitive proteases in matrix of yeast mitochondria[J]. Biochemical and Biophysical Research Communications, 1987, 144(1): 277-283.
[6] ROEDL D, TRAIDL-HOFFMANN C, RING J, et al. Serine protease inhibitor lymphoepithelial Kazal type-related inhibitor tends to be decreased in atopic dermatitis. 2009.
Using Boc-Phe-Ser-Arg-AMC to measure trypsin-like activity in stratum corneum samples[1]:
(1) Lyophilized stratum corneum samples (0.5mg) were mixed with 500μL of assay buffer (50mM Tris, 1M NaCl, 10mM EDTA, pH 8.5) and incubated at 37℃ for 1h on a shaker, then centrifuged at 13000×g for 10min at 4℃.
(2) The supernatant was obtained for protease activity assay, using Boc-Phe-Ser-Arg-AMC as the substrate for the assay of trypsin-like protease activity.
(3) Mix 100μL of supernatant sample with 100μL of assay buffer containing 0.75mM Boc-Phe-Ser-Arg-AMC, incubate at 37℃ for 5h, and measure the activity every hour.
(4) The released AMC was detected using a multi-functional microplate detector at Ex/Em of 355nm/460nm, and calibrated using AMC standards.
Using Boc-Phe-Ser-Arg-AMC to measure Kallikrein-related peptidase 6 (KLK6) activity[2]:
(1) Add 5μL of inhibitor (using 1:2 or 1:3 dilutions to achieve different final assay concentrations) or DMSO control (final assay concentration 2%) to each well of a 384-well black plate.
(2) Then add 5μL of KLK6 (final assay concentration 5nM) dissolved in reaction buffer (50mM Tris, 150mM NaCl, 1mM EDTA, 0.05% Tween-20, pH 7.5) and incubate at room temperature for 30min.
(3) Immediately before starting the kinetic measurement of fluorescence intensity (Ex, 350 ± 15nm; Em, 440 ± 20nm; every 60s for 30min), add 10μL of the substrate Boc-Phe-Ser-Arg-AMC (final assay concentration 20μM) dissolved in reaction buffer to each well.
(4) Calculate the slope of the measured signals using GraphPad Prism, normalize it relative to the DMSO control (100% activity), and then use it to generate dose-response curves.
References:
[1] GONG Z, DAI S, JIANG X, et al. Variants in KLK11, affecting signal peptide cleavage of kallikrein-related peptidase 11, cause an autosomal-dominant cornification disorder[J]. British Journal of Dermatology, 2023, 188(1): 100-111.
[2] DE VITA E, SCHÜLER P, LOVELL S, et al. Depsipeptides featuring a neutral P1 are potent inhibitors of kallikrein-related peptidase 6 with on-target cellular activity[J]. Journal of Medicinal Chemistry, 2018, 61(19): 8859-8874.
| Cas No. | 73554-90-2 | SDF | |
| Formula | C33H43N7O8 | M.Wt | 665.75 |
| 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 | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.5021 mL | 7.5103 mL | 15.0207 mL |
| 5 mM | 300.4 μL | 1.5021 mL | 3.0041 mL |
| 10 mM | 150.2 μL | 751 μL | 1.5021 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
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- Purity: >90.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 40 reference(s) in Google Scholar.)