H9c2 cells was assessed using a caspase-3 colorimetric assay (cat. no. ab39401; Abcam, Cambridge, UK), according to the manufacturer's protocols. The cell lysate was collected and incubated with 150 µM DEVD-p-nitroaniline, a fluorogenic substrate of caspase-3, at 37˚C for 2 h. The fluorescence was then measured using a clariostar Monochromator microplate reader (BMG labtech GmbH, ortenberg, Germany) with excitation at 360 nm and emission at 460 nm.
Monochromator microplate reader
Manufactured by BMG Labtech
Sourced in Germany
The Monochromator microplate reader is a versatile instrument designed for a wide range of absorbance-based assays. It utilizes monochromators to select specific wavelengths of light, enabling precise and flexible optical measurements in microplate formats.
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2 protocols using monochromator microplate reader
1
Caspase-3 Colorimetric Assay for H9c2 Cells
2
Evaluating Cancer Cell Viability with MTT Assay
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The 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine cell viability of MDA-MB-231 and MCF-7 cancer cells treated with CME. Cells were seeded in a 96-well-plate at a density of 5 × 103 cells/well. After 24 h, cells were treated with various concentrations of CME for another 24, 48, or 72 h. Cells were then treated with 0.5 mg/mL MTT (Sigma, St. Louis, MO, USA) at 37°C for 4 h. Media was removed after incubation, and DMSO (Duksan, Korea) was added to each well. Absorbance at 570 nm in each well was measured using a Clariostar Monochromator Microplate Reader (BMG LABTECH, Germany). Three independent experiments were carried out. The IC50 of CME in the cell lines were calculated using Prism 7 (GraphPad Software, CA, USA).
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