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).
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.
|Quantity:||100 x 50 μL reactions|
|Readout:||Endpoint / Kinetic|
|Label or Dye:||7-Amino-4-methylcoumarin|
|Substrate Properties:||Protein-Based Substrate|
|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).
Certificates of Analysis (COA)
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.