Cell counting kit-8 (CCK-8) assay was carried out to evaluate cell viability. Cells from each group were treated, then the cells were placed into a 96-well plate at density of 5×103/ well. After 10 μl CCK-8 reagent (Beyotime) was added to each well, the cells were incubated with CCK-8 reagent at 37° C for 2 hr. The absorbance at 450 nm was determined using a spectrophotometer.
Cck 8 reagent
Manufactured by Beyotime
Sourced in China, United States
The CCK-8 reagent is a colorimetric assay kit used to measure cell viability and proliferation. It contains a water-soluble tetrazolium salt that is reduced by living cells, producing a formazan dye that can be quantified using a spectrophotometer. The intensity of the color is directly proportional to the number of viable cells.
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Lab products found in correlation
Microplate reader, by Thermo Fisher Scientific (114 mentions)
Microplate reader, by Bio-Rad (65 mentions)
Microplate reader, by Agilent Technologies (41 mentions)
96 well plate, by Corning (11 mentions)
Microplate reader, by Molecular Devices (10 mentions)
Multiskan fc, by Thermo Fisher Scientific (10 mentions)
Model 680, by Bio-Rad (6 mentions)
Crystal violet, by Beyotime (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Infinite m200, by Tecan (5 mentions)
Synergy ht zx 22, by Agilent Technologies (4 mentions)
Fluorescence microscope, by Olympus (4 mentions)
Imark microplate reader, by Bio-Rad (4 mentions)
Microplate reader, by Tecan (4 mentions)
Cell counting kit 8 (cck8), by Beyotime (4 mentions)
Enzymatic marker, by Bio-Rad (4 mentions)
Microplate absorbance reader, by Bio-Rad (4 mentions)
Microplate reader, by Promega (4 mentions)
Prism 5, by GraphPad (4 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions)
684 protocols using cck 8 reagent
1
Cell Viability Evaluation via CCK-8
2
Cell Proliferation Assay with TSA and ITSA-1
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CNE2 and C666–1 cells were seeded into 96-well plates (4000 per well). After overnight incubation, medium was replaced with medium containing TSA with or without ITSA-1 for an additional 0, 24 and 48 h. Cells were washed with 1× PBS and then 100 μl CCK-8 reagent (Beyotime Institute of Biotechnology, Nanjing, China) and RMPI-1640 medium (CCK-8 reagent: RMPI-1640 = 1:10, v/v) was added per well. Cells were then incubated at 37 °C for 3 h. The optical density was measured using a Multiskan MKS microplate reader (Thermo Scientific, Grand Island, NY, USA) at a wavelength of 450 nm. Each experimental treatment was repeated in 6 wells, and data represent the mean ± SEM of three independent experiments.
3
Cell Viability Assay with miR-214
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LoVo cells were inoculated in a 96-well plate (3×103 cell/well) in an incubator at 37°C for 24 h. After culturing, cells were treated with PBS (control group), or were transfected with miRNA mimics (mock group) or miR-214 mimics (mimics group) for 48 h. Subsequently, 10 µl CCK-8 reagent (Beyotime Institute of Biotechnology) was added into each well, and cells were incubated with CCK-8 reagent for 4 h at 37°C. Cell viability was determined at 450 nm using a microplate absorbance spectrophotometer (Bio-Rad Laboratories, Inc.).
4
Evaluating Cell Viability in Muscle Cells
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Skeletal muscle cells were cultured in 96-well plates until 60% cell confluence. Transfection was performed with 24-h incubation. Each group had 6 parallel wells, and CCK-8 Cell viability was tested at 0 h, 24 h, 48 h, and 72 h. Specific steps of CCK-8 reagent (Beyotime, Shanghai, China,) were as follows: to each well was added 10 μL of CCK-8 reagent, and cells were incubated in 37 °C for 2 h. Finally, cells were measured by a microplate reader (OD value at 450 nm).
5
Astaxanthin Effect on Chondrocyte Viability
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According to the manufacturer’s protocol, the effect of astaxanthin on cell viability and the role of macrophages on chondrocyte viability were determined by CCK-8 assay. Chondrocytes or macrophages were seeded in 96-well plates and treated with different concentrations of astaxanthin (5, 10, 20, 40, and 80 μM) for 24 h and 48 h. Then, 100 μl of 10% CCK-8 reagent (Beyotime, China) was added to each well and incubated for 2 h, and the optical density (OD) at 450 nm of each well was measured with a microplate reader (Thermo Fisher Scientific, MA, USA).
After 7 days of coculture of macrophages and chondrocytes, CCK-8 reagent (Beyotime, China) was added to the Transwell chamber and incubated for 2 h. The cell supernatant was transferred from the Transwell chamber to a 96-well plate, and the optical density (OD) at 450 nm of each well was measured with a microplate reader (Thermo Fisher Scientific, MA, USA).
After 7 days of coculture of macrophages and chondrocytes, CCK-8 reagent (Beyotime, China) was added to the Transwell chamber and incubated for 2 h. The cell supernatant was transferred from the Transwell chamber to a 96-well plate, and the optical density (OD) at 450 nm of each well was measured with a microplate reader (Thermo Fisher Scientific, MA, USA).
6
Cell Proliferation and DDP Resistance Assay
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After transfection, cells were plated into a 96‐well plate (5 × 103 cells/well). CCK‐8 reagent (Beyotime) was added into each well at the indicated time points (0, 24, 48, and 72 h) post‐incubation. The absorbance at 450 nm was detected using a Multiskan Ascent (Thermo Fisher Scientific) to assess cell proliferation.
For drug resistance assay, DDP resistance was assessed using a CCK‐8 assay. In summary, cells were treated with different DDP concentrations for 48 h. Then, CCK‐8 reagent (Beyotime) was added to the cells and they were incubated for 2 h, followed by the determination of optical density (OD) value at 450 nm. The half inhibitory concentration (IC50) of DDP was defined as the DDP concentration when cell viability was reduced to 50%.
For drug resistance assay, DDP resistance was assessed using a CCK‐8 assay. In summary, cells were treated with different DDP concentrations for 48 h. Then, CCK‐8 reagent (Beyotime) was added to the cells and they were incubated for 2 h, followed by the determination of optical density (OD) value at 450 nm. The half inhibitory concentration (IC50) of DDP was defined as the DDP concentration when cell viability was reduced to 50%.
7
Assessing Cell Viability via CCK-8 Assay
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SV-HUC-1, T24 and EJ-1 cells were seeded in 96-well culture plates at a seeding density of 5000 cells per well, as mentioned in a previous study (24) (link). Cells were incubated with MED at concentrations of 0, 25.0, 50.0, 75.0, 100.0, 125.0, 150.0, 175.0 and 200.0 µmol L -1 for 24 hours (h) and 48 h. At a certain incubated time period (24 h or 48 h), the original complete medium was replaced with fresh complete medium with 10 % CCK-8 reagent (Beyotime Biotechnology Co., Ltd., China) and incubated for 2 hours at 37 °C. Finally, the absorbance values were obtained by Multiskan FC microplate reader (51119080, Thermo Fisher Scientific, USA) at 450 nm. The experimental results were analyzed by GraphPad Prism 8 software to calculate the IC 50 values and cell viability. The cell viability was measured by the formula presented as follows:
Cell viability (%) = [(A experimental well -A blank well )/(A control well -A blank well )] × 100 %
Cell viability (%) = [(A experimental well -A blank well )/(A control well -A blank well )] × 100 %
8
Fisetin Cytotoxicity in HEK293 Cells
9
Cell Viability Assay of HUVECs
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HUVECs in the logarithmic growth phase were detached using 0.25% trypsin to form a single-cell suspension, of which 100 μl was seeded into 96-well-plates at a concentration of 5 × 104 cells/ml. Three replicates were set in each group. Cells were incubated with 5% CO2 at 37°C for 24 and 48 h. At 1 h before the termination of culture, each well was added with 100 μl CCK-8 reagent (Beyotime). After further incubation for 1 h, the parameters of the enzyme label instrument (BD Biosciences, Franklin Lakes, NJ, USA) were set and the absorbance value of each well was measured at 450 nm.
10
Cell Viability Assay via CCK-8
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Cells were placed into 96-well plates with 5000 cells per well and cultured overnight. After addition of 10 μL of CCK-8 reagent (Beyotime, China), the cells were allowed to incubate for another 4 h at 37˚C. Afterwards, cell absorbance was detected at 450 nm by a spectrophotometer (BioRad, Hercules, CA, USA). Meanwhile, 50% inhibiting concentration (IC50) was determined from the cell viability curve.
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