A GSH-Glo Glutathione assay kit (Promega) was used to measure the total glutathione (GSH) content in the LVs following the vendor’s instructions. Briefly, 10 mg of LV heart tissue was homogenized in 1× PBS containing 2 mM EDTA, centrifuged at 12,000 rpm for 15 min at 4°C, and the supernatant was collected. A total of 50 μl of GSH-Glo Reagent was mixed with 50 μl of tissue extract (10 μg) and incubated for 30 min at RT. Next, 100 μl of luciferin detection reagent was added and incubated for an additional 15 min at RT. The signal was measured using a Turner 20/20 luminometer (Promega).
Turner 20 20 luminometer
Manufactured by Promega
The Turner 20/20 luminometer is a compact and versatile instrument designed for luminescence-based assays. It measures light output from samples, providing a quantitative analysis of luminescent signals. The luminometer's core function is to detect and measure luminescent reactions, enabling researchers to analyze various biochemical processes in their studies.
Automatically generated - may contain errors
Lab products found in correlation
Gsh glo glutathione assay kit, by Promega (2 mentions)
Dual luciferase assay system, by Promega (1 mentions)
Lipofectamine, by Thermo Fisher Scientific (1 mentions)
Lipid peroxidation assay kit, by Merck Group (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Recombinant human wnt3a protein, by R&D Systems (1 mentions)
Opti mem, by Thermo Fisher Scientific (1 mentions)
4 protocols using turner 20 20 luminometer
1
Glutathione Quantification in Cardiac Tissue
2
Measuring KRAB Domain Transcriptional Regulation
3
Glutathione and Lipid Peroxidation Assay
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Total glutathione (GSH) content in the LVs was measured by luminescence based GSH‐Glo Glutathione assay kit (Promega), and generated luminescence, corresponding to glutathione level, was detected by Turner 20/20 luminometer (Promega). Lipid peroxidation assay kit (Sigma) was used to measure malondialdehyde (MDA) level, an indicator of lipid‐peroxidation, in the LVs.
4
Assaying Wnt Pathway Activation in Ovarian Cancer
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Cells from ovarian cancer cell lines were seeded 40,000 per 50 μL in 96 well plates. Following 24 h, cells were transfected with 200 ng of TCF/LEF luciferase reporter (TOPFlash) (plasmid courtesy of Dr. Randall Moon, Upstate Biotechnology, Lake Placid, NY). Cells were transfected using Lipofectamine TM 2000 (Invitrogen, Carlsbad, CA) in Opti-MEM (Gibco/Invitrogen) per the manufacturer's instructions. After 6 h, cells were treated with 1 μM niclosamide, analog 11, or analog 32 and assayed for luciferase activity with or without Wnt3a ligand 24 h post-treatment. Luciferase activity was measured using a Turner 20/20 luminometer (Promega, Madison, WI) and was normalized to the total protein concentration as reported previously [23 (link)]. The luciferase activity was normalized to untreated control and represented as the mean ± SE for a minimum of 3 replicates. Recombinant Human Wnt-3a Protein (100 ng/ml) R&D Systems Inc. Minneapolis, MN (Catalog #5036-WN-010) was used to stimulate β-catenin driven Wnt signaling.
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