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AB112154

Proteasome 20S Activity Assay Kit (Fluorometric)

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(4 Publications)

Proteasome 20S Activity Assay Kit ab112154 is a homogeneous fluorescent assay that measures the chymotrypsin-like protease activity associated with the proteasome complex in cultured cells.
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Functional Studies - Proteasome 20S Activity Assay Kit (Fluorometric) (AB112154)
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Supplier Data

Functional Studies - Proteasome 20S Activity Assay Kit (Fluorometric) (AB112154)

Detection of Proteasome Activity in Jurkat cells. Jurkat cells were seeded on the same day at 500,000 cells/90 μL/well in a 96-well black wall/clear bottom plate. The cells were treated with or without 50 mM H2O2 for 30 minutes. The proteasome assay loading solution (100 μL/well) was added and incubated in a 5% CO2, 37 °C incubator for 3 hours. The fluorescence intensity was measured at Ex/Em = 490/525.

Key facts

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Assay type

Quantitative

Assay Platform

Microplate reader

Reactivity data

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Product details

Proteasome 20S Activity Assay Kit ab112154 is a homogeneous fluorescent assay that measures the chymotrypsin-like protease activity associated with the proteasome complex in cultured cells. ab112154 uses LLVY-R110 as a fluorogenic indicator for proteasome activities. Cleavage of LLVY-R110 by the proteasome generates strongly green fluorescent R110 that is monitored fluorometrically at 520-530 nm with excitation at 480-500 nm. The kit provides all the essential components with an optimized assay protocol. The assay is robust, and can be readily adapted for high-throughput assays to evaluate the proteasome activities or screen inhibitors in cultured cells or in solution. The assay can be performed in a convenient 96-well and 384-well fluorescence microtiter-plate format. Visit our for tips and troubleshooting.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The proteasome 20S also known as the 20S core particle is an essential component of the ubiquitin-proteasome system responsible for protein degradation. It has a mass of approximately 700 kDa and forms a barrel-shaped structure. This structure consists of four stacked rings each composed of seven subunits. The 20S proteasome is present in nearly all eukaryotic cells playing an important role in maintaining cellular protein homeostasis by degrading misfolded damaged or regulatory proteins tagged with ubiquitin.
Biological function summary

The proteasome 20S operates within a larger proteolytic complex known as the 26S proteasome essential for ATP-dependent protein degradation. The 20S core decomposes substrates into peptides through its proteolytic activity facilitated by its catalytic subunits. The proteasome activity is important for cellular processes such as cell cycle regulation apoptosis and DNA repair. This activity can be measured using a proteasome activity assay where fluorogenic assays are often used to quantify its enzymatic function.

Pathways

Proteasome 20S contributes significantly to proteostasis and the regulation of protein turnover. It involves itself in the ubiquitin-proteasome pathway which plays a pivotal role in cellular response to oxidative stress and is important for the NF-kB pathway. Its activity interacts with various proteins such as ubiquitin ligases and deubiquitinating enzymes which help target proteins for degradation. These interactions ensure timely removal of proteins preventing toxic accumulation and ensuring proper cellular functioning.

The proteasome 20S has a significant connection to neurodegenerative diseases and certain cancers. For instance in Alzheimer's disease dysregulation of proteasome activity can result in the accumulation of misfolded proteins. Similarly in multiple myeloma proteasome inhibitors are used therapeutically to reduce tumor cell survival. The interaction with proteins like tau in neurodegenerative conditions or the modulation of Bcl-2 family proteins in cancer highlight the proteasome's involvement in pathologies highlighting its potential as a therapeutic target.

Product protocols

Target data

Publications (4)

Recent publications for all applications. Explore the full list and refine your search

Methods in molecular biology (Clifton, N.J.) 2712:135-148 PubMed37578702

2023

Analysis of Protein Degradation in Ferroptosis.

Applications

Unspecified application

Species

Unspecified reactive species

Zhuojun Zhang,Lili Jiang

Human reproduction (Oxford, England) 37:2095-2112 PubMed35856882

2022

Metabolic transitions define spermatogonial stem cell maturation.

Applications

Unspecified application

Species

Unspecified reactive species

A L Voigt,R Dardari,L Su,N L M Lara,S Sinha,A Jaffer,S K Munyoki,W Alpaugh,A Dufour,J Biernaskie,K E Orwig,I Dobrinski

International journal of molecular medicine 47: PubMed33537802

2021

saponin H inhibits the proliferation of glioma cells through the A1 and A3 adenosine receptor‑mediated pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Linlin Bi,Yang Liu,Qian Yang,Xuanxuan Zhou,Hua Li,Yang Liu,Jie Li,Yunyang Lu,Haifeng Tang

FEBS open bio 6:1085-1092 PubMed27833849

2016

Rrp15 affects cell cycle, proliferation, and apoptosis in NIH3T3 cells.

Applications

Unspecified application

Species

Unspecified reactive species

Tao Wu,Mei-Xia Ren,Guo-Ping Chen,Zheng-Ming Jin,Gang Wang
View all publications
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