Cells were seeded in 96-well plates at a density of 5.0 × 103 cells/well and were then treated with the indicated concentrations of LiCl on the second day. The cells were cultured with chemotherapeutics for 24, 36 or 48 h then subjected to the MTT assay. Each sample was incubated with 20 µl of (5 mg/ml) MTT (Sigma–Aldrich; Merck KGaA) at 37 °C for 4 h. Then, the solution was discarded, and 100 μl of dimethyl sulfoxide was added. The absorbance at 495 nm due to formazan was measured by an ELISA Multiskan reader (Thermo Fisher Scientific, Inc.).
Elisa multiskan reader
Manufactured by Thermo Fisher Scientific
The ELISA Multiskan reader is a spectrophotometric plate reader used for the quantitative analysis of enzyme-linked immunosorbent assays (ELISA). The device measures the absorbance of microplate wells, which is proportional to the concentration of the analyte in the sample.
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2 protocols using elisa multiskan reader
1
MTT Cytotoxicity Assay with LiCl
2
Cisplatin Cytotoxicity on HeLa Cells
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HeLa and HeLa/Ku80-siRNA cells at the exponential phase of growth were plated in 96-well plates at a density of 1×104 cells/well and cultured in DMEM with 0, 0.5, 2, 5, 20 and 50 µg/ml cisplatin (Sigma-Aldrich; Merck KGaA) for 4 h. The medium was replaced with cisplatin-free medium. In total, ~10 µl (5 mg/ml) MTT (Sigma-Aldrich; Merck KGaA) was added to the wells when these cells were treated with cisplatin for 48 h and the plates were incubated in a 37°C incubator for 4 h. The blue formazan dye that had formed was dissolved in 150 µl dimethyl sulfoxide (Sigma-Aldrich; Merck KGaA). The absorbance (A) at 570 nm was recorded using an ELISA Multiskan reader (Thermo Fisher Scientific, Inc.). The inhibition rate was calculated as follows: Inhibition rate (%)=(1-A/A0h) ×100%. In addition, the 50% inhibitory concentration (IC50) was defined as the medicine concentration at which half of all cells were killed.
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