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Caspase glo 8 assay system

Manufactured by Promega
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The Caspase-Glo® 8 Assay System is a luminescent assay that measures the activity of caspase-8, a key enzyme involved in the initiation of the apoptotic (programmed cell death) pathway. The assay provides a quick and sensitive method for detecting caspase-8 activity in cell lysates or purified enzyme preparations.

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Lab products found in correlation

Caspase glo 3 7 assay system, by Promega (8 mentions) Caspase glo 9 assay system, by Promega (6 mentions) Propidium iodide, by Thermo Fisher Scientific (3 mentions) Cyclin b1, by Cell Signaling Technology (2 mentions) Fas associated protein with death domain fadd, by Cell Signaling Technology (2 mentions) Glyceraldehyde 3 phosphate dehydrogenase gapdh, by Santa Cruz Biotechnology (2 mentions) Kaempferol, by Merck Group (2 mentions) Dead cell apoptosis kit with annexin 5 alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Magic red caspase 3 7 kit, by Bio-Rad (1 mentions) Synergy 2 microplate reader, by Agilent Technologies (1 mentions) Dead cell apoptosis kit with annexin 5 alexa fluor 488 and propidium iodide, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Rpmi 1640 medium, by Merck Group (1 mentions) Dead cell apoptosis kit with annexin 5 fitc, by Thermo Fisher Scientific (1 mentions) Caspase glo 9, by Promega (1 mentions) Facsaria 2, by BD (1 mentions) Caspase glo 3 7, by Promega (1 mentions) Bisbenzimide h 33342 trihydrochloride hoechst 33342, by Merck Group (1 mentions)

14 protocols using caspase glo 8 assay system

1

Caspase-8 Activity Assay in HEK293T Cells

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HEK293T cells (1 × 106 cell/mL) were plated in 96-well plates and transfected with 150 ng/well indicated plasmids. The negative control group used the same quality pEGFP-N1 plasmid. The positive control group was stimulated with 2 μg/mL TNF-α for 4 h. At 20 h after transfection, the Caspase-Glo 8 assay system (Promega) was tested using SpectraMax i3 according to the manufacturer's instructions (scanning at full wavelength).
Hou J., Pang Y, & Li Q. (2020). Comprehensive Evolutionary Analysis of Lamprey TNFR-Associated Factors (TRAFs) and Receptor-Interacting Protein Kinase (RIPKs) and Insights Into the Functional Characterization of TRAF3/6 and RIPK1. Frontiers in Immunology, 11, 663.
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2

Caspase Activity Measurement Protocol

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The activity of caspase-3, caspase-8 and caspase-9 was analyzed using Caspase-Glo®3/7 Assay System, Caspase-Glo®8 Assay System and Caspase-Glo®9 Assay System (Promega, Madison, WI, USA) according to the manufacturer's instructions. Briefly, 96-well plates containing cells were removed from the incubator and the plates were allowed to equilibrate to room temperature. Caspase-Glo® reagent (100 µl) was added to each well of a white-walled 96-well plate containing 100 µl of blank or cells in culture medium. The contents of the wells were gently mixed and incubated at room temperature for 30 min. Finally, the luminescence of each sample was assessed on a plate-reading luminometer (PerkinElemer EnVision; PerkinElemer, Waltham, MA, USA) as directed by the manufacturer's instructions.
Lin Z., Liu W., Xiao C., Fan Y., Zhuang G, & Qi Z. (2018). TIPE2 inhibits GC via regulation of cell proliferation, apoptosis and inflammation. Oncology Reports, 40(3), 1307-1316.
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3

Measuring Caspase Activity in MEFs

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MEFs were seeded in 96-well plates and stimulated as described. A luciferase-based caspase assay was used to measure caspases-3/7 (Caspase-Glo® 3/7 Assay Systems, Promega) and caspase-8 (Caspase-Glo 8 Assay System, Promega) activity following stimulation according to manufacturer's protocol. Samples were read on a Victor3 1420 multilabel automated plate reader, and triplicates were performed for each condition (means ± S.D. are presented). The luciferase assays in HEK293T cells have been described previously (3 (link)), and the human NLRX1 promoter (−1515;+1)-driven luciferase construct was purchased from GeneCopoeia.
Soares F., Tattoli I., Rahman M.A., Robertson S.J., Belcheva A., Liu D., Streutker C., Winer S., Winer D.A., Martin A., Philpott D.J., Arnoult D, & Girardin S.E. (2014). The Mitochondrial Protein NLRX1 Controls the Balance between Extrinsic and Intrinsic Apoptosis. The Journal of Biological Chemistry, 289(28), 19317-19330.
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4

Caspase Activation Assay for TRAIL

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Cells were treated GST-TRAIL or drozibumab as indicated in the text and figure legends. For caspase inhibition, cells were preincubated with 100 uM ZVAD-FMK or DMSO for two hours and subsequently incubated overnight with 2500 ng/ml drozitumab or 2500 ng/ml drozitumab plus 10 μg/ml F(ab’)2. Caspase activity was assessed using the Caspase-Glo® 3/7 assay system (Cat # G8092, Promega Corporation, Madison, WI) and/or Caspase-Glo® 8 assay system (Cat # G8202, Promega Corporation, Madison, WI) as previously described [27 (link)]. Three independent experiments normalized to the control cells +/− SE were performed.
Dine J.L., O’Sullivan C.C., Voeller D., Greer Y.E., Chavez K.J., Conway C.M., Sinclair S., Stone B., Amiri-Kordestani L., Merchant A.S., Hewitt S.M., Steinberg S.M., Swain S.M, & Lipkowitz S. (2016). The TRAIL Receptor Agonist Drozitumab Targets Basal B Triple Negative Breast Cancer Cells that Express Vimentin and Axl. Breast cancer research and treatment, 155(2), 235-251.
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5

Evaluating FasL-induced Apoptosis in Oral Cancer Cells

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OSCC‐derived cells (SJGs) and OK cells were seeded on Corning® Rat Tail Collagen I (Corning, NY, USA) coated 96‐well plates (30,000 cells/well). The next day, cells were treated for 6 h with 0–400 ng/ml human FasL (Sigma‐Aldrich). Measurements of caspase activity were performed using the Caspase‐Glo® 3/7 Assay System (Promega, Madison, WI, USA) and the Caspase‐Glo® 8 Assay System (Promega), following the manufacturer's instructions.
Bhosale P.G., Kennedy R.A, & Watt F.M. (2023). Caspase activation in tumour‐infiltrating lymphocytes is associated with lymph node metastasis in oral squamous cell carcinoma. The Journal of Pathology, 261(1), 43-54.
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6

Kaempferol Induces Apoptosis in Cancer Cells

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Kaempferol was purchased from Sigma. Dead Cell Apoptosis Kit with Annexin V Alexa Fluor® 488 and propidium iodide (PI) was purchased from ThermoFisher Scientific (Waltham, MA, USA). Caspase-Glo® 3/7 Assay Systems, Caspase-Glo® 8 Assay Systems and Caspase-Glo® 9 Assay Systems were purchased from Promega (Madison, WI, USA). Antibodies against p21 Waf1/Cip1 (p21), p-Cdc25C (Ser 216), Cdc25C, p-Cdc2 (Tyr 15), Cdc2, Cyclin B1, p53, death receptor 5 (DR5), Fas, and Fas-Associated protein with Death Domain (FADD) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Antibodies against phosphor-checkpoint kinase 2 (Chk2) (Thr68), Chk2, poly [ADP-ribose] polymerase 1 (PARP-1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).
Gao Y., Yin J., Rankin G.O, & Chen Y.C. (2018). Kaempferol Induces G2/M Cell Cycle Arrest via Checkpoint Kinase 2 and Promotes Apoptosis via Death Receptors in Human Ovarian Carcinoma A2780/CP70 Cells. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(5), 1095.
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7

Apoptosis Assay in MB Cells

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To measure apoptosis in the MB cell lines, we used the Caspase-Glo® 8 Assay Systems (#G8200, Promega). We plated 1,000 cells/well in 200 μL of MEM media and each sample was plated in triplicate. The specified drugs were incubated with the cells for 24 hours and then apoptotic levels were measured using a plate reader to determine the luminescence readings according to the manufacturer’s directions.
Nitta R.T., Bolin S., Luo E., Solow-Codero D.E., Samghabadi P., Purzner T., Aujla P.S., Nwagbo G., Cho Y.J, & Li G. (2019). Casein Kinase 2 inhibition sensitizes medulloblastoma to temozolomide. Oncogene, 38(42), 6867-6879.
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8

Apoptosis Assay in MB Cells

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To measure apoptosis in the MB cell lines, we used the Caspase-Glo® 8 Assay Systems (#G8200, Promega). We plated 1,000 cells/well in 200 μL of MEM media and each sample was plated in triplicate. The specified drugs were incubated with the cells for 24 hours and then apoptotic levels were measured using a plate reader to determine the luminescence readings according to the manufacturer’s directions.
Nitta R.T., Bolin S., Luo E., Solow-Codero D.E., Samghabadi P., Purzner T., Aujla P.S., Nwagbo G., Cho Y.J, & Li G. (2019). Casein Kinase 2 inhibition sensitizes medulloblastoma to temozolomide. Oncogene, 38(42), 6867-6879.
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9

Caspase-8 and -9 Activity Assay

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Activities of caspase-8 and -9 were measured using Caspase-Glo® 8 Assay Systems and Caspase-Glo® 9 Assay Systems (Promega, Madison, WI, USA) according to manufacturer’s protocol. Luminescent signal which was proportional to caspase activity was measured an hour after the addition of Caspase-Glo® Reagent to the post-treated cells in 1:1 ratio.
Yap H.Y., Tan N.H., Ng S.T., Tan C.S, & Fung S.Y. (2018). Molecular attributes and apoptosis-inducing activities of a putative serine protease isolated from Tiger Milk mushroom (Lignosus rhinocerus) sclerotium against breast cancer cells in vitro. PeerJ, 6, e4940.
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10

Caspase 8 and Caspase 3 Assay

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Caspase 8 activity was measured using the Caspase-Glo® 8 Assay Systems (Promega) according to manufacturer’s instructions. Caspase 3 activity was measured using the Magic Red Caspase 3/7 Kit (BioRad) according to manufacturer’s instructions.
Tölken L.A., Paulikat A.D., Jachmann L.H., Reder A., Salazar M.G., Medina L.M., Michalik S., Völker U., Svensson M., Norrby-Teglund A., Hoff K.J., Lammers M, & Siemens N. (2024). Reduced interleukin-18 secretion by human monocytic cells in response to infections with hyper-virulent Streptococcus pyogenes. Journal of Biomedical Science, 31, 26.
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