Ac-Trp-Leu-Ala-AMC (WLA-AMC)

Catalog number: SBB-PS0008, 2 mg

Ac-WLA-AMC is a fluorogenic peptidyl substrate for measuring chymotrypsin-like activity of the proteasome. Hydrolysis of this substrate by the β5 subunit of the 20S proteasome is monitored by observing fluorescence at an Excitation wavelength of 345nm and Emission at 445nm. [1][2][3]

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Ac-WLA-AMC (Acetyl-Trp-Leu-Ala-AMC) is a 7-amino-4-methylcoumarin labeled fluorogenic peptidyl substrate hydrolyzed by the β5 subunit of the 20S proteasome. Chymotrypsin-like activity can be measured using a working concentration of 20-50μM substrate. This substrate is specific to the constitutive proteasome, and is not hydrolyzed efficiently by the immunoproteasome. Cleavage of this peptide by the proteasome or other enzymes liberates the fluorophore AMC causing a strong fluorescent signal which is detected at an Excitation wavelength of 345nm and Emission wavelength of 445nm. 20S Proteasome enzyme requires activation with 0.035% SDS in the assay buffer.

For Research Use Only, Not For Use In Humans.


Product Overview
Quantity: 2.0 mg
Molecular Weight: 587.7 Da
Purity: >99% by HPLC
Readout: Endpoint / Kinetic
Label or Dye: 7-Amino-4-methylcoumarin
Detection Method: Fluorescence
Substrate Properties: Protein-Based Substrate, C32H37N5O6
Excitation/Emission (nm): 345/445
Storage Buffer: Lyophilized
Storage Store at 4°C after product arrival. After preparing a stock in DMSO ( ≥10 mM) store product at -20°C to −80°C. It is recommended to make multiple aliquots after the first thaw to ensure best performance.

Figures & Data


Figure 1. Ac-WLA-AMC, Chemical Structure.
Structure of Ac-WLA-AMC.

20S Proteasome Activity

Figure 2. 20S Proteasome vs. Immunoproteasome Activity.
WLA-AMC exhibits a high specific activity and preference for 20S Proteasome compared to Immunoproteasome.

Certificates of Analysis (COA)

Citations & References

1) Ettari, Roberta, et al. "Immunoproteasome-selective inhibitors: a promising strategy to treat hematologic malignancies, autoimmune and inflammatory diseases." Current medicinal chemistry 23.12 (2016): 1217-1238.

2) Miller, Zachary, Wooin Lee, and Kyung Bo Kim. "The immunoproteasome as a therapeutic target for hematological malignancies." Current cancer drug targets 14.6 (2014): 537-548.

3) Blackburn, Christopher, et al. "Characterization of a new series of non-covalent proteasome inhibitors with exquisite potency and selectivity for the 20S β5-subunit." Biochemical journal 430.3 (2010): 461-476.