20S Immunoproteasome Activity Assay Kit (Fluorometric)

A plate-based fluorometric assay to measure the activity of the 20S immunoproteasome.
 
MILPITAS, Calif. - Dec. 10, 2020 - PRLog -- Proteasome mediated protein degradation is one of the major pathways for the degradation of damaged, misfolded or aggregated proteins in cells. The 26S proteasome is the most common form of the proteasome that degrades all ubiquitinated proteins. It is an ATP-dependent proteolytic complex containing a barrel-shaped core (20S proteasome) and two caps (19S regulatory complexes) at both ends.

The 20S immunoproteasome is a specialized form of the proteasome, which is expressed during oxidative stress or by stimulation from proinflammatory cytokines. During oxidative stress and inflammatory processes, the catalytic subunits β1, β2 and β5 of the 20S proteasome core are replaced by the inducible subunits β1i/LMP2, β2i/MECL1 and β5i/LMP7, respectively. This allows the 20S immunoproteasome to cleave proteins differently as compared to the 20S proteasome due to a shift towards chymotrypsin-like catalytic activity. Additionally, these inducible catalytic subunits play an important role in the generation of new antigenic peptides for presentation via the major histocompatibility class I pathway.

BioVision's 20S Immunoproteasome Activity Assay Kit uses a plate-based fluorometric assay to measure the activity of the 20S immunoproteasome. In this assay, the 20S immunoproteasome cleaves a fluorogenic substrate to release 7-amino-4-methylcoumarin, a fluorescent compound that produces a stable signal at Ex/Em = 345/445 nm. The kit can detect as low as 0.02 µU (0.02 pmol substrate proteolysis per minute) of 20S immunoproteasome under assay conditions.

Figures. (a). AMC Standard Curve. (b). Enzyme kinetics using mouse spleen lysate (94 µg protein/well) and J774A.1 cell lysate (25 µg protein/well) (c). Specific activity of 20S Immunoproteasome in mouse spleen lysate and J774A.1 cell lysate. Experiments were conducted according to the kit protocol.

For more information on this Assay Kit Cat# K2051, Visit: https://bit.ly/3qURuUO

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