Catalog number: SBB-PS0009, 2 mg
Ac-ANW-AMC is a fluorogenic peptidyl substrate for measuring chymotrypsin-like activity of the immunoproteasome. Hydrolysis of this substrate by the β5i subunit of the immunoproteasome is monitored by observing fluorescence at an Excitation wavelength of 345nm and Emission at 445nm. 
Ac-ANW-AMC (Acetyl-Ala-Asn-Trp-AMC) is a 7-amino-4-methylcoumarin labeled fluorogenic peptidyl substrate hydrolyzed by the β5i subunit of the 20S immunoproteasome. Chymotrypsin-like activity can be measured using a working concentration of 20-50μM substrate. This substrate is specific to the immunoproteasome, and is not hydrolyzed efficiently by the constitutive proteasome. Cleavage of this peptide by the immunoproteasome 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.
|Molecular Weight:||588.6 Da|
|Purity:||>99% by HPLC|
|Readout:||Endpoint / Kinetic|
|Label or Dye:||7-Amino-4-methylcoumarin|
|Substrate Properties:||Protein-Based Substrate, C30H32N6O7|
|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-ANW-AMC, Chemical Structure.
Structure of Ac-ANW-AMC.
Figure 2. 20S Immunoproteasome vs. 20S Constitutive Proteasome Activity.
ANW-AMC exhibits a specificity for 20S Immunoproteasome compared to 20S constitutive proteasome.
Certificates of Analysis (COA)
Citations & References
1) Singh, Pradeep K., et al. "Immunoproteasome β5i‐Selective Dipeptidomimetic Inhibitors." ChemMedChem 11.19 (2016): 2127-2131.
2) De Groot, Karina A., et al. "Pharmacodynamic monitoring of (immuno) proteasome inhibition during bortezomib treatment of a critically ill patient with lupus nephritis and myocarditis." Lupus science & medicine 2.1 (2015): e000121.
3) Cornish Carmony, Kimberly, et al. "Elucidating the Catalytic Subunit Composition of Distinct Proteasome Subtypes: A Crosslinking Approach Employing Bifunctional Activity‐Based Probes." ChemBioChem 16.2 (2015): 284-292.