Cell counting kit 8 cck 8 reagent

After 24-hour transfection, the cells were digested and counted to prepare cell
suspension. A 96-well plate was added into 100 µL cell suspension (the
concentration was 1 × 103 cells/well). Cells were cultured under standard
conditions. When testing the proliferation, 10 µL Cell Counting Kit-8 (CCK8)
reagent (Beijing Solarbio Science & Technology, Co, Ltd) was put into each
well. The cells were cultured for 1.5 hours at 37 °C in a CO₂incubator, and the cell proliferation was determined every 24 hours. The
OD₄₅₀ was detected utilizing a microplate reader.

Cell proliferation was measured using Cell Counting Kit-8 (CCK-8) reagent (Solarbio, Beijing, China) according to the manufacturer’s protocol. The N2a cells were seeded into 96-well plates (5 × 10³ cells per well) and incubated. Upon reaching 40% confluency, the cells were transfected with miRIDIAN miR-380-3p mimic or hairpin inhibitor. The corresponding scrambled sequences were adopted in negative control groups. The transfected cells were then exposed to 0, 50, 100, or 200 µM of PQ for 48 h, followed by cell proliferation assessment. A mixture of 90 μl medium and 10 μl CCK-8 solution was used to replace the PQ-stained medium in each well, incubated for 2 h and the absorbance values were measured at 450 nm using a microplate reader.

Cells were seeded into 96-well plates at a density of 1×10⁴ cells/well at a temperature of 37 °C and a carbon dioxide content of 5%. Zeaxanthin was added to the cells at concentrations of 1, 3, 10, 30, and 100 μM for 24 h, or they were treated with zeaxanthin (17 μM, the IC₅₀ value) for 3, 6, 12, 24, and 36 h. Ten micro-litres of Cell Counting Kit-8 (CCK-8) reagent (Solarbio) was added and after 2 h, and samples were examined using a microplate reader (BioTek, Instruments Inc., Winooski, VT, USA). Optical density (OD) values were recorded at 450 nm to determine the percentage of viable cells. The percentage of cell viability was calculated at 50% inhibitory concentration (IC₅₀) using Graphpad Prism 5 software (GraphPad Software, Inc., La Jolla, CA, USA).28 (link) All experiments and measurements were performed in triplicate.

EJ and SW780 cells were seeded into 96-well plates and then treated with different concentrations of desloratadine (0, 0.5, 1, 2, 4, 8, 16, 24, 32, and 64 µM, MedChemExpress) for 24 hours. After treatment, Cell Counting Kit 8 (CCK8) reagent (10 μL/well, Beijing Solarbio Science & Technology) was added into each well, followed by incubation at 37 °C for 90 minutes. The optical density (OD) was measured at 450 nm.

MHCC97H cells were seeded at 2×10³ cells/well and incubated at 37°C for 12 h in a 96-well plate. Prior to detection, Cell Counting Kit-8 (CCK-8) Reagent (10 µl/well; Solarbio Science & Technology Co., Ltd.) was added to each well and cells were incubated for 2 h at 37°C in a 5% CO₂ atmosphere. The absorbance was then measured at 450 nm using a microplate reader and the OD value of each well was used to represent cell proliferation.