Transfected A2780 and SKOV3 cells were reseeded onto 96-well plates. At the indicated time points, 10 μL cell counting kit 8 (CCK8) reagent (Roche) was added to each well. After cultured for 4 h, the optical density (OD) value was detected at 450 nm using a microplate reader.
Cck 8 reagent
Manufactured by Roche
Sourced in Switzerland
The CCK-8 reagent is a colorimetric assay kit used to measure cell viability and proliferation in cell-based studies. It contains a water-soluble tetrazolium salt that is reduced by living cells, producing a colored formazan dye that can be quantified using a spectrophotometer.
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8 protocols using cck 8 reagent
1
Cell Proliferation Assay with CCK8
2
Doxorubicin-induced H9C2 cell cytotoxicity
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H9C2 cells were seeded (0.5x103 cells/ml) into 96-well plates and incubated with various concentrations of DOX (0, 0.1, 1 and 10 µM) for 12 and 24 h at 37˚C (0 µM as control group). Subsequently, 10 µl CCK-8 reagent (Roche Diagnostics) was added to each well according to the manufacturer's protocol and incubated for 2 h at 37˚C. The optical density of each well was measured at a wavelength of 490 nm using a microplate reader.
To confirm the effect of catalpol on H9C2 cells, H9C2 cells were cultured with various concentrations of catalpol (0, 10, 20, 40 and 80 µmol/l) for 24 h at 37˚C (0 µM as control group). Then, to confirm the effect of DOX-induced H9C2 cardiomyocytes, catalpol administration in each dose group (10, 20, 40 and 80 µmol/l) after H9C2 cells treatment with DOX (1 µM) for 24 h at 37˚C, respectively. According to the aforementioned method, 10 µl CCK-8 regent was added to each well in accordance with the manufacturer's protocol and incubated for 2 h at 37˚C. The optical density of each well was measured at a wavelength of 490 nm using a microplate reader.
To confirm the effect of catalpol on H9C2 cells, H9C2 cells were cultured with various concentrations of catalpol (0, 10, 20, 40 and 80 µmol/l) for 24 h at 37˚C (0 µM as control group). Then, to confirm the effect of DOX-induced H9C2 cardiomyocytes, catalpol administration in each dose group (10, 20, 40 and 80 µmol/l) after H9C2 cells treatment with DOX (1 µM) for 24 h at 37˚C, respectively. According to the aforementioned method, 10 µl CCK-8 regent was added to each well in accordance with the manufacturer's protocol and incubated for 2 h at 37˚C. The optical density of each well was measured at a wavelength of 490 nm using a microplate reader.
3
Cell Viability Assay with PAB Treatment
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After human TE-1, ECA-109, KYSE410, and KYSE520 cells were treated with PAB at various concentrations, cells were seeded in 96-well plates at a density of 1x103 cells/mL and incubated at 24, 48, and 72 hours. Subsequently, 10 μL CCK-8 reagent (Roche Diagnostics, Basel, Switzerland) was added into each single well. Following 1 hour of incubation, the absorbance of CCK-8 was measured by an automated plate reader at 490 nm (Bio-Rad, Hercules, CA, USA).
4
Cell Viability Assessment of MC3T3-E1 Cells
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At 4 8h after transfection, MC3T3-E1 cells were seeded in 96-well plates at a density of 0.5×103 cells/ml and incubated for at 1, 3, and 5 days. Then, 10 μl of CCK-8 reagent (Roche Diagnostics, Basel, Switzerland) was added into every well. After 2 h of incubation, the optical density was measured at 490 nm using a microplate reader.
5
Evaluating CD4+ T Cell Proliferation
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To assess cell proliferation, we stained CD4+ T cells with CFSE Cell Division Tracker Kit (BioLegend, San Diego, CA) in accordance with the manufacturer's instructions. After activation and co-culture with sorted mregDCs or non-mregDCs, CD4+ T cells were harvested on day 4 and evaluated using flow cytometry. Cell proliferation was also quantified using CCK-8 reagent (Roche, Mannheim, Germany) in line with the manufacturer's specifications. The results were assayed by applying ELx808 absorbance readers (BioTek Instruments, Winooski, VT).
6
Cell Viability Assay in 96-well Plates
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SCL-1 and A431 cell lines were cultured in 96-well plates at a density of 5×103 cells/well at 37°C for 72 h. Cell Counting Kit-8 (CCK-8) reagent (10 μL; Roche Diagnostics, Germany) was then added to each well, and the cells were incubated for 2 h. Absorbance was measured at 450 nm using a Thermo FC Microplate Reader (Molecular Devices, Biobase, Japan).
7
Cell Viability Assay for Breast Cancer
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Breast cancer cells seeded in triplicate in microtiter plates (96 wells) with a density of 3 × 103 cells per well in 100-μl medium were treated as indicated in the text and figure legends for 72 h. The CCK-8 reagent (Roche, Shanghai, China) was added to cells at a concentration of 10 μL/well in 100 μL culture medium and the samples were incubated at 37°C for 2–4 hours. The optical density (OD) of each sample was measured using a microplate reader at 450 nM. The percentages of absorbance relative to those of untreated control samples were plotted as a linear function of drug concentration. Inhibition of cell viability was measured by percentage of viable cells relative to the control: % inhibition = 100% × ODT/ODC, where ODT is the average OD value of the treated samples and ODC is the average OD value of the control samples. Combination index (CI) was calculated by an equation reported in a previous report [24 (link)]. CI less than 0.9 indicates synergism; 0.9 to 1.1, additivity; and greater than 1.1, antagonism.
8
Cell Proliferation Assay with CCK8
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Cell proliferation was measured using CCK8 reagent (Roche) according to the manufacturer’s instructions. Cells were seeded into 96-well plates at a density of 3000 cells/well. When cells were cultured for the indicated time, CCK8 reagent was added. All experiments were performed in triplicate.
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