Product nameProteasome Activity Assay Kit
Sample typeCell Lysate
Assay typeEnzyme activity
Assay time1h 00m
Species reactivityReacts with: Other species, Mammals
Proteasome Activity Assay Kit (ab107921) takes advantage of the chymotrypsin-like activity, using an AMC-tagged peptide substrate (Proteasome Substrate (Succ-LLVY-AMC in DMSO), which releases free, highly 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
The 20S proteasome assembly is the functional protease structure with chymotrypsin-like, trypsin-like and caspase-like protease activities
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.
Storage instructionsStore at -20°C. Please refer to protocols.
Storage bufferPreservative: None
Components Identifier 100 tests AMC Standard (1mM in DMSO) Yellow 1 x 100µl Positive Control (lyophilized) Green 1 vial Proteasome Substrate (in DMSO) Red 1 x 100µl Proteasome Assay Buffer WM 1 x 25ml Proteasome Inhibitor (in DMSO) Blue 1 x 100µl
Standard curve: mean of duplicates (+/- SD) with background reads subtracted
Proteasome Activity measured in Jurkat lysate
Proteasome Activity measured in HeLa lysate
Proteasome Activity measured in cell lysates showing activity (mU) per 1 mln cells.
Samples with the concentration of 3.3e6 cells/mL were used. 50 μl of each sample was used.
ab107921 has been referenced in 27 publications.
- Merlos Rodrigo MA et al. Proteomic Signature of Neuroblastoma Cells UKF-NB-4 Reveals Key Role of Lysosomal Sequestration and the Proteasome Complex in Acquiring Chemoresistance to Cisplatin. J Proteome Res 18:1255-1263 (2019). PubMed: 30592607
- Bono F et al. Nicotine prevents alpha-synuclein accumulation in mouse and human iPSC-derived dopaminergic neurons through activation of the dopamine D3- acetylcholine nicotinic receptor heteromer. Neurobiol Dis 129:1-12 (2019). PubMed: 31051233
- Wang C et al. Methyltransferase-like 21e inhibits 26S proteasome activity to facilitate hypertrophy of type IIb myofibers. FASEB J 33:9672-9684 (2019). PubMed: 31162944
- McMillan EA et al. A Genome-wide Functional Signature Ontology Map and Applications to Natural Product Mechanism of Action Discovery. Cell Chem Biol 26:1380-1392.e6 (2019). PubMed: 31378711
- Leskelä S et al. C9orf72 Proteins Regulate Autophagy and Undergo Autophagosomal or Proteasomal Degradation in a Cell Type-Dependent Manner. Cells 8:N/A (2019). PubMed: 31658762