Cell counting kit-8 (CCK-8) assay was used to assess cell proliferation. Cells were attached to 96-well plate at indicated times and cell viability was measured by adding CCK8 reagents (Dalian Meilun Biology Technology Co., Ltd.) with incubation for 2 h. The OD450 value of each sample was quantified using a spectrophotometer. Cell proliferation was also evaluated by a colony formation assay. MDA-MB-231 cells were plated at a low density (2×103 cells/well) in 6-well plate, and culture media with 30% serum (Every Green; Zhejiang Tianhang Biotechnology Co., Ltd. was replaced every 3 days. Following 2 weeks of incubation, 4% paraformaldehyde (Dalian Meilun Biology Technology Co., Ltd.), was used to fix the cells at room temperature for 30 min. After staining with 0.1% crystal violet (Servicebio) for 15 min at room temperature, excess crystal violet was washed away with PBS, and the plaques were imaged and analyzed using the ImageJ software (version 1.54f; National Institutes of Health).
Cck8 reagents
Manufactured by Meilun
Sourced in China
CCK8 reagents are a colorimetric assay used to measure cell viability and proliferation. The reagents contain a tetrazolium salt that is reduced by living cells, resulting in a colored product that can be measured spectrophotometrically.
Automatically generated - may contain errors
2 protocols using cck8 reagents
1
Cell Proliferation Assays for Cancer Research
2
Cell Viability Assay of PA-Treated Min6 Cells
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CCK-8 assay was performed to determine the cell viability under PA treatment and the protective activity of the tested compounds in PA-treated Min6 cells. In brief, Min6 cells were seeded in 96-well plates and cultured in DMEM overnight. Min6 cells were treated with tested compounds at indicated concentration and PA (300 μmol/L) for 24 h. After washing cells with PBS, fresh DMEM and CCK-8 reagents (Meilunbio, Dalian, China) were added and cells were incubated for 2 h in a 37 °C-incubator containing 5% CO2. Absorbance at 450 nm was read on a microplate reader (Molecular Devices, SpectraMax 190, USA).
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