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Proteasome glo

Manufactured by Promega
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Proteasome-Glo is a luminescent-based assay used to measure proteasome activity in cell lysates or purified proteasome samples. The assay utilizes a cell-based substrate that is cleaved by the proteasome, resulting in the generation of a luminescent signal proportional to proteasome activity.

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Lab products found in correlation

Np 40, by Merck Group (1 mentions) Softmax pro 5 microplate reader, by Molecular Devices (1 mentions) Mg132, by Enzo Life Sciences (1 mentions) Caspase glo 3 7 assay, by Promega (1 mentions) Ros glo h2o2 assay, by Promega (1 mentions) Proteasome glo chymotrypsin like cell based assay, by Promega (1 mentions) Spectramax m2 reader, by Molecular Devices (1 mentions) Proteasome glo assay, by Promega (1 mentions) Prestoblue, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Proteasome-GloTM Chymotrypsin-like Cell-Based Assays kit Proteasome GloTM Assay Proteasome-GloTM Assay Systems Proteasome-GloTM Assay kit Proteasome-Glo Chymotrypsin-like Cell-Based Assay kit Proteasome-Glo™ Assay System Proteasome-Glo Chymotrypsin-Like Assay kit Proteasome-Glo assay Proteasome-Glo 3-substrate cell-based assay system Proteasome-Glo™ Reagent

11 protocols using proteasome glo

1

Proteasome Activity in MCF7 Cells

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MCF7 cells were plated in triplicates in 96 well plates at a density of 20,000 cells per well. 24 hours later, chymotrypsin-like activity of the proteasome was assayed, using the Proteasome-Glo™ assay (Promega), according to manufactures protocol. The activity levels were normalized to the relative cell number that was measured using the fluorescent detection of resazurin dye reduction (544-nm excitation and 590-nm emission).
Ben-David U., Siranosian B., Ha G., Tang H., Oren Y., Hinohara K., Strathdee C.A., Dempster J., Lyons N.J., Burns R., Nag A., Kugener G., Cimini B., Tsvetkov P., Maruvka Y.E., O’Rourke R., Garrity A., Tubelli A.A., Bandopadhayay P., Tsherniak A., Vazquez F., Wong B., Birger C., Ghandi M., Thorner A.R., Bittker J.A., Meyerson M., Getz G., Beroukhim R, & Golub T.R. (2018). Genetic and transcriptional evolution alters cancer cell line drug response. Nature, 560(7718), 325-330.
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2

Proteasome Activity Assays in Cells

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Trypsin-like proteasome activity was measured in a live-cell assay using a luminogenic substrate (Z-LRR-aminoluciferin; Proteasome-Glo, Promega Corporation Madison, WI). Briefly, to cells released into a suspension (10,000 cells/100 μL) 100 μL of Proteasome-Glo cell-based reagent was added. After mixing and incubating (6 min, room temperature) luminescence was measured using the SoftMax Pro 5 microplate reader (Molecular Devices, Inc. Sunnyvale, CA). Samples were assayed in duplicate.
Chymotrypsin-like proteasome activity was measured using a fluorescence assay that employs an AMC-tagged peptide substrate (Succ-LLVY-AMC; BioVision Incorporated, Mountain View, CA) and cells homogenized in 0.5 % NP-40 (Sigma-Aldrich Corp., St. Louis, MO). Fluorescence (excitation/emission, 350/440 nm) was measured in the microplate reader (37 °C from 30–60 min). Nonspecific (non-proteasome) fluorescence measured in the presence of a proteasome inhibitor was subtracted, values were adjusted to protein concentration and proteasome activity was determined from a standard curve of AMC fluorescence. Samples were assayed in duplicate.
Sparrow J.R., Zhou J., Ghosh S.K, & Liu Z. (2014). Bisretinoid Degradation and the Ubiquitin-Proteasome System. Advances in experimental medicine and biology, 801, 593-600.
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3

Proteasome Activity Assay Protocol

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Proteasome activity was evaluated using Proteasome-Glo (Chymotrypsin-like/Caspase-like/Trypsin-like) cell-based assays (Promega).47 (link) Cells were treated with the indicated concentrations of MG132 and the luminescence signal was quantified by using an Orion Microplate Luminometer. Cell viability was measured in parallel to exclude any bias. Data were normalized to the control and the specific proteasome activities indicated in percent.
Cerella C., Muller F., Gaigneaux A., Radogna F., Viry E., Chateauvieux S., Dicato M, & Diederich M. (2015). Early downregulation of Mcl-1 regulates apoptosis triggered by cardiac glycoside UNBS1450. Cell Death & Disease, 6(6), e1782-.
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4

Proteasome Activity Measurement Protocol

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We measured the cell’s ability to cleave Suc-LLVY-aminoluciferin utilizing Proteasome-Glo (Promega) following a one-hour treatment with the noted proteasome inhibitor and measured luminescence. Results shown are from at least two biological replicates.
Hong A.L., Tseng Y.Y., Wala J.A., Kim W.J., Kynnap B.D., Doshi M.B., Kugener G., Sandoval G.J., Howard T.P., Li J., Yang X., Tillgren M., Ghandi M., Sayeed A., Deasy R., Ward A., McSteen B., Labella K.M., Keskula P., Tracy A., Connor C., Clinton C.M., Church A.J., Crompton B.D., Janeway K.A., Van Hare B., Sandak D., Gjoerup O., Bandopadhayay P., Clemons P.A., Schreiber S.L., Root D.E., Gokhale P.C., Chi S.N., Mullen E.A., Roberts C.W., Kadoch C., Beroukhim R., Ligon K.L., Boehm J.S, & Hahn W.C. (2019). Renal medullary carcinomas depend upon SMARCB1 loss and are sensitive to proteasome inhibition. eLife, 8, e44161.
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5

Proteasome Activity Assay in BV-2 Cells

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1 × 104 BV-2 cells were plated in a clear bottom, black 96-well plate. 24 h following, they were treated with IFNγ in the absence and presence of MG-132 (10 μM, Enzo Life Sciences) for 24 h then subject to a live cell chymotrypsin-like proteasome activity assay (Proteasome-Glo, Promega) per manufacturer instructions.
Moritz K.E., McCormack N.M., Abera M.B., Viollet C., Yauger Y.J., Sukumar G., Dalgard C.L, & Burnett B.G. (2017). The role of the immunoproteasome in interferon-γ-mediated microglial activation. Scientific Reports, 7, 9365.
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6

Proteasome Activity Assay in MPNST Cells

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MPNST cells were seeded at 104 cells/well in 96-well plates in triplicate and treated with bortezomib. Chymotrypsin-like activity of the 20S proteasome was determined according to the manufactory’s instructions (Proteasome-Glo, Promega). Luminescence was read in Victor-3 automated plate reader (Perkim-Elmer).
Yamashita A., Baia G., Ho J., Velarde E., Wong J., Gallia G., Belzberg A., Kimura E, & Riggins G. (2014). PRECLINICAL EVALUATION OF THE COMBINATION OF mTOR AND PROTEASOME INHIBITORS WITH RADIOTHERAPY IN MALIGNANT PERIPHERAL NERVE SHEATH TUMORS. Journal of neuro-oncology, 118(1), 83-92.
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7

Proteasome and Caspase Activity Assay

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Cells were seeded in a white 96-well plate at an 8000 cells/well density and incubated overnight for attachment. Cells were treated with either DMSO control, MG-132 (1 µM), staurosporin (0.1 μM) or Foldlin (at IC50 concentration) for a total incubation time of 8 h (proteasome) or 16 h (caspase test). Proteasomal activity and caspase activity were monitored according to the manufacturer’s guidelines (ROS-Glo H2O2 assay, Promega, Leiden, The Netherlands; Proteasome-Glo, Promega, Leiden, The Netherlands, caspase-GLO 3/7 assay, Promega, Leiden, The Netherlands).
Naus E., Derweduwe M., Lampi Y., Claeys A., Pauwels J., Langenberg T., Claes F., Xu J., Haemels V., Atak Z.K., van der Kant R., Van Durme J., De Baets G., Ligon K.L., Fiers M., Gevaert K., Aerts S., Rousseau F., Schymkowitz J, & De Smet F. (2023). Reduced Levels of Misfolded and Aggregated Mutant p53 by Proteostatic Activation. Cells, 12(6), 960.
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8

Measuring Proteasome Chymotrypsin-like Activity

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A cell-based luminescent proteasome assay (Proteasome-Glo, Promega, Madison, WI, USA) was used to measure chymotrypsin-like proteasome activity as described [51 (link)]. The assay contains the luminogenic peptide substrate Suc-LLVY-aminoluciferin for determination of chymotrypsin-like activity of the proteasome.
Keskin I., Forsgren E., Lehmann M., Andersen P.M., Brännström T., Lange D.J., Synofzik M., Nordström U., Zetterström P., Marklund S.L, & Gilthorpe J.D. (2019). The molecular pathogenesis of superoxide dismutase 1-linked ALS is promoted by low oxygen tension. Acta Neuropathologica, 138(1), 85-101.
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9

Proteasome Activity in MCF7 Cells

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MCF7 cells were plated in triplicates in 96 well plates at a density of 20,000 cells per well. 24 hours later, chymotrypsin-like activity of the proteasome was assayed, using the Proteasome-Glo™ assay (Promega), according to manufactures protocol. The activity levels were normalized to the relative cell number that was measured using the fluorescent detection of resazurin dye reduction (544-nm excitation and 590-nm emission).
Ben-David U., Siranosian B., Ha G., Tang H., Oren Y., Hinohara K., Strathdee C.A., Dempster J., Lyons N.J., Burns R., Nag A., Kugener G., Cimini B., Tsvetkov P., Maruvka Y.E., O’Rourke R., Garrity A., Tubelli A.A., Bandopadhayay P., Tsherniak A., Vazquez F., Wong B., Birger C., Ghandi M., Thorner A.R., Bittker J.A., Meyerson M., Getz G., Beroukhim R, & Golub T.R. (2018). Genetic and transcriptional evolution alters cancer cell line drug response. Nature, 560(7718), 325-330.
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10

Proteasomal Activity in Ire1α MEFs

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Proteasomal activity of wild-type MEFs and Ire1α knockout (Ire1α−/−) MEFs were determined by Proteasome-Glo assay (Promega) following the manufacturer’s instructions.
Kanekura K., Ma X., Murphy J.T., Zhu L.J., Diwan A, & Urano F. (2015). IRE1 prevents endoplasmic reticulum membrane permeabilization and cell death under pathological conditions. Science signaling, 8(382), ra62.
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