Description
Key facts
Detection method:Fluorescent,
Sample types:Cell Lysate,
Assay type:Enzyme activity,
Assay time:1h,
Assay Platform:Microplate reader
Product details:
Proteasome Activity Assay Kit ab107921 is a simple and reliable assay to quantify proteasome activity.
Proteasome activity assay principle
The 20S proteasome assembly is the functional protease structure in the proteasome, with chymotrypsin-like, trypsin-like and caspase-like protease activities. The assay is based on measuring the chymotrypsin-like activity.
The assay uses an AMC-tagged peptide substrate (Proteasome Substrate (Succ-LLVY-AMC in DMSO), which releases free, fluorescent AMC (Ex/Em 350/440 nm) in the presence of proteolytic activity.
The kit also includes a positive control (Jurkat cell lysate with significant proteasome activity), and a specific proteasome inhibitor MG-132 which suppresses all proteolytic activity due to proteasomes. This permits differentiation of proteasome activity from other protease activity which may be present in samples.
Proteasome activity assay protocol summary
- add samples, standards and positive control to wells
- add proteasome inhibitor to half of sample and positive control wells
- add proteasome substrate to sample and control wells
- incubate for 20 min
- analyze with microplate reader, incubate for further 30 min, analyze again
How other researchers are using Proteasome Activity Assay Kit ab107921
ab107921 has been used in a variety of sample types including:
- Mouse ovarian surface epithelial cell lines (1)
- Mouse intestine samples (2)
- Mouse brain lysates (3)
References: 1-Tailor D et al. 2023; 2-Hurley M et al. 2023; 3-Vulin M et al. 2023
Related and recommended products
Proteasome 20S Activity Assay Kit (Fluorometric)
ab112154
uses an alternative fluorophore and a similar principle to this assay, with an LLVY proteasome substrate.
This product was previously called K245 Biovision Proteasome Activity Fluorometric Assay Kit. Biovision was acquired by Abcam in 2021.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
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.
Proteasomes are large protein complexes responsible for degrading ubiquitinated proteins into peptides. This process is essential for maintaining protein homeostasis within the cell. The proteasome often referred to as the 26S proteasome consists of a core particle with a mass of roughly 700 kDa and a regulatory particle. They are expressed in the cytoplasm and nucleus of eukaryotic cells where they perform the bulk of cellular protein degradation. Other names for the proteasome include multicatalytic proteinase complex and prosome.
Biological function summary
The ubiquitin-proteasome system orchestrates the controlled degradation of unneeded or damaged proteins regulating numerous cellular mechanisms. The proteasome forms a complex with ubiquitin chains tagged to substrate proteins guiding them for degradation. Proteasomal activity ensures turnover of proteins involved in cell cycle progression signal transduction and immune responses. By controlling protein levels proteasome function influences many biological outcomes and maintains cellular equilibrium.
Pathways
The proteasome's function integrates deeply into the ubiquitin-proteasome pathway critical for protein catabolism and cellular control processes. Another significant pathway is the regulation of apoptosis where the proteasome controls the levels of proteins like p53 an important regulator of cell death. In these pathways the proteasome collaborates with other proteins such as ubiquitin ligases playing role in fine-tuning the cellular processes involved in survival and programmed cell death.
Proteasome dysfunction contributes to neurodegenerative diseases like Parkinson's and Alzheimer's. Disruption in proteasomal activity leads to aggregation of damaged proteins which can have pathological consequences. Cancer is another area where proteasomes play an important role; certain cancers show dysregulated proteasomal activity resulting in abnormal degradation of tumor suppressor proteins like p53. Understanding proteasome markers helps in diagnosing and evaluating these diseases linking proteasomes to both prognosis and potential therapeutic interventions.
