20S Immunoproteasome Kit, human PBMC

Catalog number: SBB-KP0037

This kit is designed to test for specific activity of 20S immunoproteasome. The kit provides purified 20S immunoproteasome and is designed to test for Chymotrypsin-like activity (Suc-LLVY-AMC), and Caspase-like activity of the immunoproteasome subunits ß1i/ PSMB9 (PAL-AMC), and ß5i/PSMB8 (ANW-AMC).[1][2][3][4][5][6]

In stock
SKU
0037
$375.00

Description

This kit is designed to test for specific activity of 20S immunoproteasome. The kit provides purified 20S immunoproteasome and is designed to test for Chymotrypsin-like activity (Suc-LLVY-AMC), and Caspase-like activity of the immunoproteasome subunits ß1i/ PSMB9 (PAL-AMC), and ß5i/PSMB8 (ANW-AMC). Additionally, we have included the compound, ONX-0914, which can be used to inhibit specifically the subunit ß5i/LMP7 20S immunoproteasome. All peptide substrates are conjugated to AMC, which upon proteasome catalyzed hydrolyses display fluorescence at Excitation = 345 nm, Emission = 445 nm; allowing for a real-time read out of 20S immunoproteasome specific activity.

For Research Use Only, Not For Use In Humans.

Specifications

Product Overview
Quantity: 100 x 50 μL reactions
Purity: > 95%
Readout: Endpoint / Kinetic
Label or Dye: 7-Amino-4-methylcoumarin
Detection Method: Fluorescence
Substrate Properties: Protein-Based Substrate
Excitation/Emission (nm): 345/445
Storage -80C, Avoid multiple freeze / thaw cycles. It is recommended to make aliquots of each reaction component upon first time use.

Figures & Data

20S Immunoproteasome Activity

Figure 1. 20S Immunoproteasome Activity Raw Data Output: Several wells of Immunoproteasome shown digesting LLVY, PAL, and ANW-AMC over time +/- 1x (40μM) inhibitor (ONX-0914).

Citations & References

1) Singh, Pradeep K., et al. “Immunoproteasome ß5iSelective 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.
4) Park, Ji Eun, et al. “PSMB9 codon 60 polymorphisms have no impact on the activity of the immunoproteasome catalytic subunit B1i expressed in multiple types of solid cancer.” PloS one 8.9 (2013): e73732.
5) Miller, Zachary, et al. “Inhibitors of the immunoproteasome: current status and future directions.” Current pharmaceutical design 19.22 (2013): 4140-4151.
6) Dubiella, Christian. Development and Characterization of Selective Immunoproteasome Inhibitors. Diss. Universität München, 2015.