RealTime-Glo MT assay (Promega GmbH, Walldorf, Germany) was performed to examine the impact of CSE on the proliferation of metabolically active cells. The 16HBE cells were first seeded in different densities (5 × 103, 104 and 2 × 104 cells/well) in the 96-well white-walled and transparent bottom plate (Greiner Bio-One GmbH, Frickenhausen, Germany) to achieve an optimal cell density in which the signal remains linear throughout the experiment. Next, the cells were seeded at 104 and incubated with 2X RealTime-Glo reagent. The luminescence at assay start (0 h) was then measured with a microplate reader (BioTek HT, BioTek Instrument GmbH, Bad Friedrichshall, Germany). In the next rounds, following incubating of the cells with the reagent the cells were stimulated with medium control or different concentrations of CSE (5–20%) and incubated in at 37 °C with 7.5% CO2. The luminescence was measured at 1 h, 3 h, 7 h, and 19 h post-treatment in the plate. The luminescent signal was plotted versus cell number using a linear curve fit in GraphPad prism version 9.3.1.
Realtime glo mt assay
Manufactured by Promega
Sourced in Germany, United Kingdom, Italy
The RealTime-Glo MT Cell Viability Assay is a luminescent assay that measures the metabolic activity of cells in real-time. It uses a cell-permeant pro-luciferin substrate and a proprietary thermostable luciferase enzyme to provide a quantitative readout of cell metabolism.
Automatically generated - may contain errors
4 protocols using realtime glo mt assay
1
Evaluating Cell Proliferation with RealTime-Glo MT Assay
2
Evaluating Pioglitazone and UK 5099 Effects
3
Compatibility of Integra Bilayer with hASCs
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Integra bilayer was incubated in petri dish supplemented with culture medium (see below) for 24 h at 37°C in order to test its compatibility with hASCs. Petri dish was then observed by an inverted phase-contrast microscope Zeiss Axiovert 200 (Arese, MI, Italy) connected to an AxioCam ICm1camera (Zeiss); photographs were acquired by the Axiovision Rel 4.8 software (Zeiss).
Cell viability was determined by the RealTime-Glo™ MT assay (Promega, Milano, Italy) according to the manufacturer's instructions. Briefly, 200 µl of hASCs suspension (containing about 1 × 103 cells) were seeded, together with the scaffold and the assay reagent, into 24-well assay plates and cultivated for 6 days at 37°C in 5% of CO2. Every 24 h luminescent signal has been recorded using the Infinite F200 plate reader (Tecan Group, Männedorf, Switzerland). The experiments were performed in triplicate and normalized compared with cell viability in absence of the scaffold. For all the experiments we have used hASCs at the passage 5.
Cell viability was determined by the RealTime-Glo™ MT assay (Promega, Milano, Italy) according to the manufacturer's instructions. Briefly, 200 µl of hASCs suspension (containing about 1 × 103 cells) were seeded, together with the scaffold and the assay reagent, into 24-well assay plates and cultivated for 6 days at 37°C in 5% of CO2. Every 24 h luminescent signal has been recorded using the Infinite F200 plate reader (Tecan Group, Männedorf, Switzerland). The experiments were performed in triplicate and normalized compared with cell viability in absence of the scaffold. For all the experiments we have used hASCs at the passage 5.
4
Determining Apoptotic Effects of BCL-2 Antagonists
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ABT-737 was the selective BCL-2/BCL-XL antagonist (Active Biochem Ltd.), A-1210477 was the MCL-1-selective antagonist (MedChemExpress) were solubilized in DMSO at different concentrations (0.1, 1.0, 5.0 and 10.0 µM). HL60, MOLM13, MV4-11 and OCI-AML3 cell lines were treated for 72 h at 37°C with A-1210477 and/or ABT-737. DMSO was used as a control at a concentration of 0.001%. A Real-Time-Glo™ MT assay (Promega Corporation) was used to assess the cell viability according to the manufacturer's protocol. The present study also used a fluorescence microscope to visualize the cell luminescence.
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