Apoptosis and cell viability studies were performed using the Caspase-Glo and CellTiter-Glo luminescence-based assays (Promega) [41 (link)]. Cells were plated in 96-well flat-bottomed plates (Perkin Elmer) after transfection and incubated for 48 hours (Caspase-Glo) or 72 hours (CellTiter-Glo). Luminescence was measured with a BioTek Synergy 2 Luminometer (BioTek). Data were normalized to the cells transfected with non-targeting control siRNA.
Biotek synergy 2 luminometer
Manufactured by Agilent Technologies
Sourced in United States
The BioTek Synergy 2 luminometer is a compact and versatile instrument designed for various luminescence-based assays. It features a sensitive photomultiplier tube detector and provides accurate and reproducible results for a wide range of applications.
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Lab products found in correlation
4 protocols using biotek synergy 2 luminometer
1
Apoptosis and Cell Viability Assays
2
Mandarin Fish Brain Total RNA Extraction
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Total RNA of mandarin fish brains was extracted with Trizol Reagent (TaKaRa, Tokyo, Japan) according to the manual. The purity and quantity of total RNA were determined using the BioTek Synergy 2 luminometer (BioTek, Winooski, VT, USA), and integrity of total RNA was checked using electrophoresis in 2% agarose gel (Biowest Agarose, Madrid, Spain). The cDNAs were obtained from 1 μg total RNA with the Revert Aid™ Reverse Transcriptase (TaKaRa, Tokyo, Japan) according to the manufacturer’s instructions.
3
Automated Bacterial Luciferase Assay
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Cells were grown in LB to mid-log phase and diluted 1:100 in fresh LB. Dilutions were plated in quadruplet (250 μl each) in a 96-well polystyrene Costar plate (white with a clear bottom; Fisher Scientific, USA). The luciferase activity of each strain was measured on a BioTek Synergy 2 luminometer (BioTek, USA) with continuous slow shaking at 30°C. Luciferase luminescence was measured at a sensitivity setting of 200, and culture optical density was measured at 600 nm every 10 minutes for 24 hours. Final luciferase activity values were calculated by normalizing luciferase luminescence to culture density. Data shown represents the average of at least four biological replicates.
4
ATP Luminescence Assay in T84 Cells
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T84 cells were plated in 96 well plates (50 000 cells/well) and incubated overnight in DMEM/F12 medium supplemented with 2 mM L-glutamine, 8% foetal bovine serum and 1% penicillin/streptomycin (PEST). The next day, medium was discarded and the cells were washed twice with Krebs-Ringer Hepes (KRH) buffer (120 mM NaCl, 4.7 mM KCl, 2.2 mM CaCl2, 10 mM HEPES, 0.12 mM KH2PO4, 0.12 mM MgSO4 in milliQ deionised water, pH 7.4). The buffer was removed and the cells were incubated with 100 µL of KRH or KRH with 50 mM K+, both with 0.1% fatty acid-free BSA, containing SR101 (10 µM), carbenoxolone (30 µM), mefloquine (30 µM) or vehicle control (DMSO, maximum assay concentration 0.05%) for 15 minutes. ATP luminescence was measured using the CellTitre Glo (ref G7570) kit from Promega (Madison, WI 53711 USA). Briefly, aliquots (50 µL) of medium were then transferred to a new plate and 50 µL of the reagent were added before the plate was read on a BioTek Synergy 2 luminometer (BioTek, Winooski, VT, USA). Reagent (50 µL) was also added to the cells and luminescence was read following lysis. Luminescence in the medium was normalized to luminescence in the cells.
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