Ac-Ala-Asn-Trp-AMC (ANW-AMC)

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. [1][2][3]

In stock
SKU
0009
$109.00

Description

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.

Specifications

Product Overview
Quantity: 2.0 mg
Molecular Weight: 588.6 Da
Purity: >99% by HPLC
Readout: Endpoint / Kinetic
Label or Dye: 7-Amino-4-methylcoumarin
Detection Method: Fluorescence
Substrate Properties: Protein-Based Substrate, C30H32N6O7
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

Ac-ANW-AMC

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

Immunoproteasome Activity

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.