20S Proteasome Kit, human RBC

Catalog number: SBB-KP0038

This kit is designed to test for specific activity of 20S proteasome. The kit provides purified 20S proteasome and is designed to test for Chymotrypsin-like activity (Suc-LLVY-AMC), Caspase-like activity of the proteasome subunits ß1/PSMB6 (LLE-AMC), and Chymotrypsin-like activity of ß5/PSMB5 (WLA-AMC). [1][2][3][4]

In stock
SKU
0038
$295.00

Description

This kit is designed to test for specific activity of 20S proteasome. The kit provides purified 20S proteasome and is designed to test for Chymotrypsin-like activity (Suc-LLVY-AMC), Caspase-like activity of the proteasome subunits ß1/PSMB6 (LLE-AMC), and Chymotrypsin-like activity of ß5/PSMB5 (WLA-AMC). Additionally we have included the compound, MG-132, which can be used to inhibit the proteasome. 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 proteasome 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 Proteasome Activity

Figure 1. 20S Proteasome Activity Raw Data Output: Several wells of Proteasome shown digesting LLVY, WLA, and LLE-AMC over time +/- 1x (40uM) inhibitor (MG-132).

Certificates of Analysis (COA)

Citations & References

1) Smith DM, Chang SC, Park S, Finley D, Cheng Y, Goldberg AL (Sep 2007). “Docking of the proteasomal ATPases’ carboxyl termini in the 20S proteasome’s alpha ring opens the gate for substrate entry”. Molecular Cell. 27 (5): 731–44. doi:10.1016/j.molcel.2007.06.033. PMC 2083707Freely accessible. PMID 17803938.
2) Wilk S, Orlowski M (Mar 1983). “Evidence that pituitary cation-sensitive neutral endopeptidase is a multicatalytic protease complex”. Journal of Neurochemistry. 40 (3): 842–9. doi:10.1111/j.1471-4159.1983.tb08056.x. PMID 6338156.
3) Kisselev, Alexei F., and Alfred L. Goldberg. “Monitoring activity and inhibition of 26S proteasomes with fluorogenic peptide substrates.” Methods in enzymology 398 (2005): 364-378.
4) 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.