Cells were cultured in a 96-well plate at the density of 1×104 cells/well and subjected to plasmid transfection or C. burnetii infection. At the appointed time points, the cells were detected using a proteasome activity detection kit (Promega, G8660). Briefly, the assay buffer was prepared by dissolving luciferin detection reagent into the Proteasome-Glo buffer. Then, the Suc-LLVY-Glo substrate was added at the ratio of 1:200 to generate the reaction reagent. After 30 min of incubation at room temperature, 100 μl of the reaction reagent were added to the cells, and the luminescence of each sample was measured in a plate reading luminometer after an incubation for a minimum of 10 minutes.
Proteasome activity detection kit
Manufactured by Promega
The Proteasome Activity Detection Kit is a laboratory tool designed to measure the activity of proteasomes, which are large protein complexes responsible for the degradation of unwanted or damaged proteins within cells. This kit provides a quantitative assay to assess the proteolytic function of proteasomes in samples, enabling researchers to study their role in cellular processes and protein homeostasis.
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Lab products found in correlation
Anti β5i, by Abcam (1 mentions)
Pr 957, by Selleck Chemicals (1 mentions)
Anti β5, by Abcam (1 mentions)
Annexin 5 pi apoptosis detection kit, by BD (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Anti β actin, by Abcam (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Bortezomib, by Selleck Chemicals (1 mentions)
Mg132, by Selleck Chemicals (1 mentions)
2 protocols using proteasome activity detection kit
1
Proteasome Activity Quantification
2
Proteasome Inhibition Assay in Cells
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Corilagin analytical standard (purity, >99%) was purchased from Sigma-Aldrich (Merck KGaA) and TMZ from Selleck Chemicals. Anti-β1, anti-β2, anti-β5, anti-β1i, anti-β2i and anti-β5i antibodies were procured from Abcam; anti-β-actin and horseradish peroxidase (HRP)-conjugated goat anti-mouse and anti-rabbit IgG secondary antibodies were acquired from OriGene Technologies, Inc.; MG-132, bortezomib, carfizomib and PR-957 were procured from Selleck Chemicals. DMEM, streptomycin penicillin, and fetal calf serum were obtained from HyClone (GE Healthcare Life Sciences); trypsin was obtained from Gibco (Thermo Fisher Scientific, Inc.). BCA protein assay kit was purchased from Beyotime Institute of Biotechnology, an Annexin V/PI apoptosis detection kit from BD Biosciences and proteasome activity detection kit from Promega Corporation.
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