(Suc-LLVY)2R110
(Suc-LLVY)2R110 is a sentive fluorogenic substrate for 20S proteasome, calpains and other chymotrypsin-like proteases. The non-fluorescent substrate generates a bright green fluorescent rhodamine 110 product that has an emission spectra that can be easily detected with a FITC filter set. This rhodamine 110 substrate is much more sensitive than the AMC-, AFC- or 4-nitroaniline-based substrates. The most common form of the proteasome is known as the 26S proteasome that contains one 20S core particle structure and two 19S regulatory caps. All 20S particles consist of four stacked heptameric ring structures that are themselves composed of two different types of subunits; alpha subunits are structural in nature, whereas beta subunits are predominantly catalytic. The outer two rings in the stack consist of seven alpha subunits each, which serve as docking domains for the regulatory particles and the alpha subunits N-termini form a gate that blocks unregulated access of substrates to the interior cavity. The inner two rings each consist of seven beta subunits and contain the protease active sites that perform the proteolysis reactions.
Example protocol
AT A GLANCE
Important notes
Following protocol only provides a guideline, and should be modified according to your specific needs.
Product Number |
Indicators |
Excitation |
Emission |
13451 |
(Suc-LLVY)2R110 |
498 nm |
520 nm |
13453 |
Suc-LLVY-AMC |
351 nm |
430 nm |
13455 |
(Ac-ANW)2R110 |
498 nm |
520 nm |
13465 |
(Ac-KQL)2R110 |
498 nm |
520 nm |
13466 |
(Z-LLE)2R110 |
498 nm |
520 nm |
13467 |
(Ac-PAL)2R110 |
498 nm |
520 nm |
13468 |
(Ac-WLA)2R110 |
498 nm |
520 nm |
PREPARATION OF STOCK SOLUTION
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
1. (Suc-LLVY)2R110 stock solution:
Prepare a 5 to 10 mM stock solution in DMSO.
PREPARATION OF WORKING SOLUTION
Component | Volume |
Substrate stock solution (10 mM) | 25 µL |
DTT (1M) | 100 µL |
EDTA (100 mM) | 400 µL |
Hepes Buffer (25 mM), pH = 7.4 | 10 mL |
Total volume | 10.53 mL |
SAMPLE EXPERIMENTAL PROTOCOL
- Mix equal volume of the protesome standards or samples with 2X fluorescent proteasome substrate assay solution, and incubate the solutions at room temperature for at least 1 hour.
- Monitor the fluorescence using fluorescent microplate readers at 498/520 nm.
Spectrum
Open in Advanced Spectrum Viewer
Product family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) |
Suc-LLVY-Aminoluciferin | 362 | 499 | 18000 |
Suc-LLVY-AMC | 341 | 441 | - |
Citations
View all 5 citations: Citation Explorer
Cytotoxic unsaturated electrophilic compounds commonly target the ubiquitin proteasome system
Authors: Selvaraju, Karthik and Mofers, Arjan and Pellegrini, Paola and Salomonsson, Johannes and Ahlner, Alex and ra , undefined and Morad, Vivian and Hillert, Ellin-Kristina and Espinosa, Belen and Arnér, Elias SJ and Jensen, Lasse and others, undefined
Journal: Scientific Reports (2019): 9841
Authors: Selvaraju, Karthik and Mofers, Arjan and Pellegrini, Paola and Salomonsson, Johannes and Ahlner, Alex and ra , undefined and Morad, Vivian and Hillert, Ellin-Kristina and Espinosa, Belen and Arnér, Elias SJ and Jensen, Lasse and others, undefined
Journal: Scientific Reports (2019): 9841
Diabetogenic agent alloxan is a proteasome inhibitor
Authors: Zhou, Wenjuan and Wei, Lingling and Xiao, Ting and Lai, Chunyou and Peng, Min and Xu, Lingli and Luo, Xiangwei and Deng, Shaoping and Zhang, Fengxue
Journal: Biochemical and Biophysical Research Communications (2017): 400--406
Authors: Zhou, Wenjuan and Wei, Lingling and Xiao, Ting and Lai, Chunyou and Peng, Min and Xu, Lingli and Luo, Xiangwei and Deng, Shaoping and Zhang, Fengxue
Journal: Biochemical and Biophysical Research Communications (2017): 400--406
Delineation of molecular pathways involved in cardiomyopathies caused by troponin T mutations
Authors: Gilda, Jennifer E and Lai, Xianyin and Witzmann, Frank A and Gomes, Aldrin V
Journal: Molecular & Cellular Proteomics (2016): 1962--1981
Authors: Gilda, Jennifer E and Lai, Xianyin and Witzmann, Frank A and Gomes, Aldrin V
Journal: Molecular & Cellular Proteomics (2016): 1962--1981
Diclofenac induces proteasome and mitochondrial dysfunction in murine cardiomyocytes and hearts
Authors: Ghosh, Rajeshwary and Goswami, Sumanta K and Feitoza, Luis Felipe BB and Hammock, Bruce and Gomes, Aldrin V
Journal: International Journal of Cardiology (2016): 923--935
Authors: Ghosh, Rajeshwary and Goswami, Sumanta K and Feitoza, Luis Felipe BB and Hammock, Bruce and Gomes, Aldrin V
Journal: International Journal of Cardiology (2016): 923--935
Advanced-glycation-end-product-induced formation of immunoproteasomes: involvement of RAGE and Jak2/STAT1
Authors: Grimm, Stefanie and Ott, Christiane and Hörlacher, Melanie and Weber, Daniela and Höhn, Annika and Grune, Tilman
Journal: Biochemical Journal (2012): 127--139
Authors: Grimm, Stefanie and Ott, Christiane and Hörlacher, Melanie and Weber, Daniela and Höhn, Annika and Grune, Tilman
Journal: Biochemical Journal (2012): 127--139
References
View all 36 references: Citation Explorer
Alveolar extracellular 20S proteasome in patients with acute respiratory distress syndrome
Authors: Sixt SU, Adamzik M, Spyrka D, Saul B, Hakenbeck J, Wohlschlaeger J, Costabel U, Kloss A, Giesebrecht J, Dahlmann B, Peters J.
Journal: Am J Respir Crit Care Med (2009): 1098
Authors: Sixt SU, Adamzik M, Spyrka D, Saul B, Hakenbeck J, Wohlschlaeger J, Costabel U, Kloss A, Giesebrecht J, Dahlmann B, Peters J.
Journal: Am J Respir Crit Care Med (2009): 1098
Quinone reductase acts as a redox switch of the 20S yeast proteasome
Authors: Sollner S, Schober M, Wagner A, Prem A, Lorkova L, Palfey BA, Groll M, Macheroux P.
Journal: EMBO Rep (2009): 65
Authors: Sollner S, Schober M, Wagner A, Prem A, Lorkova L, Palfey BA, Groll M, Macheroux P.
Journal: EMBO Rep (2009): 65
Comparative expression analysis and characterization of 20S proteasomes in human intestinal tissues: The proteasome pattern as diagnostic tool for IBD patients
Authors: Visekruna A, Joeris T, Schmidt N, Lawrenz M, Ritz JP, Buhr HJ, Steinhoff U.
Journal: Inflamm Bowel Dis (2009): 526
Authors: Visekruna A, Joeris T, Schmidt N, Lawrenz M, Ritz JP, Buhr HJ, Steinhoff U.
Journal: Inflamm Bowel Dis (2009): 526
Enzymatic properties of the 20S proteasome in wheat endosperm and its biochemical characteristics after seed imbibition
Authors: Shi C, Rui Q, Xu LL.
Journal: Plant Biol (Stuttg) (2009): 849
Authors: Shi C, Rui Q, Xu LL.
Journal: Plant Biol (Stuttg) (2009): 849
BCL-2 family regulation by the 20S proteasome inhibitor bortezomib
Authors: Fennell DA, Chacko A, Mutti L.
Journal: Oncogene (2008): 1189
Authors: Fennell DA, Chacko A, Mutti L.
Journal: Oncogene (2008): 1189
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