Water peroxide concentrations were determined in gPE (1 mmol/L) and cPE (1 mmol/L) standards using the Red hydrogen peroxide assay kit (Enzo Life Science; Lörrach, Germany) following the manufacturer´s protocol. Briefly, gPE and cPE were dissolved in the assay buffer and transferred into a 96-well plate. A solution composed of red peroxidase substrate and 0.8 U/mL peroxidase was added and fluorescence (λexc=540 nm, λem=590 nm) was recorded using the Paradigm™ Detection Platform (Molecular devices, Salzburg, Austria).
Paradigm detection platform
Manufactured by Molecular Devices
Sourced in Austria
The Paradigm™ Detection Platform is a versatile detection system designed for a range of applications in life sciences research. It provides reliable and accurate measurements of various assays, including absorbance, fluorescence, and luminescence. The Paradigm™ platform is capable of analyzing multiple samples simultaneously, offering efficient and high-throughput data collection.
Automatically generated - may contain errors
Lab products found in correlation
Cellstar, by Greiner (2 mentions)
Red hydrogen peroxide assay kit, by Enzo Life Sciences (1 mentions)
Cleaved caspase 3 antibody, by Cell Signaling Technology (1 mentions)
Gefitinib, by Cell Signaling Technology (1 mentions)
Crizotinib, by Cell Signaling Technology (1 mentions)
Alamar blue assay kit, by Thermo Fisher Scientific (1 mentions)
Gefitinib, by LC Laboratories (1 mentions)
Cisplatin, by Merck Group (1 mentions)
Paclitaxel, by LC Laboratories (1 mentions)
Trametinib, by LC Laboratories (1 mentions)
Crizotinib, by LC Laboratories (1 mentions)
Erlotinib, by LC Laboratories (1 mentions)
Caspase glo 3 7 assay, by Promega (1 mentions)
4 protocols using paradigm detection platform
1
Quantifying Peroxide Levels in Organic Compounds
2
Antiproliferative Effects of Kinase Inhibitors
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Gefitinib (#G-4408), erlotinib (#E-4007), trametinib (#T-8123), crizotinib (#C-7900) and paclitaxel (#P-9600) were purchased from LC Laboratories (Woburn, MA, USA). Cisplatin (#479306) was purchased from Sigma Aldrich (St. Louis, MO, USA). 2D and 3D cultures were incubated in the presence of the drugs for 72 hours. Cell viability was determined using the commercially available alamarBlue® assay kit (Thermo Fisher Scientific, Madison, WI, USA). Apoptosis was analyzed using the Caspase-Glo® 3/7 assay multiplexed with the MultiTox-Fluor GF-AFC life stain (Promega, Madison, WI, USA). Luminescence and fluorescence intensities were measured using the Paradigm detection platform (Molecular Devices, Sunnyvale, CA, USA). Statistical analysis was done using the GraphPad Prism Software 7.03 (GraphPad Software Inc., La Jolla, CA, USA). For the in situ analyses of apoptosis in cocultures the cells were incubated in the presence of 50 nM Gefitinib or 20 nM crizotinib for 24 hours and processed for immunofluorescent microscopy using the cleaved caspase-3 antibody (Cell Signaling Technology, #9661) that specifically detects apoptotic cells.
3
Paraquat-Induced Oxidative Stress Assay
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Cells were grown on flat bottom black 96-well plates (Greiner CELLSTAR®) overnight in DMEM/Ham's F12 medium without FBS. Next day the medium was replaced with transparent DMEM containing DCFDA (10 µmol/L) and incubated (37 °C, 1 h). Cells were washed and incubated in DMEM. After 1 h the medium was changed to DMEM containing different concentrations of PQ and the fluorescence (485/535 nm) was recorded for 3 h on a Paradigm™ Detection Platform (Molecular devices, Salzburg, Austria).
4
Fluorescent Oxidative Stress Assay
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Cells were grown on a flat bottom black 96-well plates (Greiner CELLSTAR®). Following FA-loading, medium was replaced with transparent DMEM containing DCFDA (Sigma Aldrich GmbH, Germany; 10 µmol/L) and incubated (37 °C, 1 h). Cells were washed and incubated in DMEM. After 30 min the medium was exchanged to DMEM with or without CoCl2 and the fluorescence (485/535 nm) was recorded for 5 h on a Paradigm™ Detection Platform (Molecular devices, Salzburg, Austria).
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