After treatment for 48 h, 10% of CCK-8 reagent (Yeasen, Shanghai, China) was added to each well and incubated for further 4 h at 37°C. The metabolic activation of CCK-8 was quantified by measuring the absorbance at 450 nm by using the spectrophotometer (Thermo Fisher Scientific Oy, Vantaa, Finland).
Cck 8 reagent
Manufactured by Yeasen
Sourced in China, United States
The CCK-8 reagent is a colorimetric assay kit used to measure cell viability and cytotoxicity. It utilizes a water-soluble tetrazolium salt that is reduced by living cells, resulting in the formation of a water-soluble formazan dye. The intensity of the color produced is proportional to the number of viable cells, allowing for the quantification of cell proliferation and cytotoxicity.
Automatically generated - may contain errors
Lab products found in correlation
Microplate reader, by Thermo Fisher Scientific (10 mentions)
Microplate reader, by Agilent Technologies (7 mentions)
Multiskan sky, by Thermo Fisher Scientific (3 mentions)
Varioskan lux, by Thermo Fisher Scientific (2 mentions)
Crystal violet, by Beyotime (2 mentions)
Fluorescence microscope, by Olympus (2 mentions)
Multiskan go, by Thermo Fisher Scientific (2 mentions)
Facsverse, by BD (1 mentions)
Cell light edu apollo567 in vitro kit, by RiboBio (1 mentions)
Cycletest plus dna reagent kit, by BD (1 mentions)
Synergy 2, by Agilent Technologies (1 mentions)
Prism v8, by GraphPad (1 mentions)
Microplate spectrophotometer, by Agilent Technologies (1 mentions)
Compusyn software, by ComboSyn (1 mentions)
Synergy 4, by Agilent Technologies (1 mentions)
Edu cell proliferation detection kit, by Beyotime (1 mentions)
Imark microplate absorbance reader, by Bio-Rad (1 mentions)
Microplate reader, by Molecular Devices (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Nano 100, by Allsheng (1 mentions)
78 protocols using cck 8 reagent
1
Cell Viability Assay with CCK-8
2
Evaluating EIF4E3 Effects on Osteosarcoma
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Osteosarcoma cells were transfected with the human EIF4E3 overexpression plasmid and incubated in serum-free, antibiotic-free medium for 4 to 6 h before replacing the complete medium. Following this, cell viability was assessed by a colony formation assay in which 1 × 10 3 osteosarcoma cells were inoculated in 6-well plates and cultured for 12 days before staining with crystal violet. For the CCK-8 assay, 5 × 103 transfected osteosarcoma cells were incorporated into a 96-well culture plate for 24 h. Afterward, 10 μL of CCK-8 reagent (Yeasen, China) was added to measure the absorbance at 450 nm. Transwell assays were conducted to analyze the effect of EIF4E3 on osteosarcoma cell migration and invasion. In the upper chamber, osteosarcoma cells were inoculated at 5 × 104/well into serum-free medium, and in the lower chamber, 500 μL of medium containing 10% FBS was added. Cells were seeded into the upper chamber with precoated Matrigel For invasion. One day later, cells migrating to the subsurface of the PET membrane were immobilized with 4% paraformaldehyde and stained with crystal violet.
3
Cell Proliferation and Cell Cycle Analysis
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Following the cell transfection in a 96-well for 24 h, 10 μL of CCK-8 reagent (Yeasen, Shanghai, China) was added into each well and incubated for 1.5 h in a 37 °C incubator. Finally, the Varioskan LUX (Thermo Scientific, Singapore) was used to measure the absorbance at 450 nm. Cell proliferation was detected using a Cell-Light EdU Apollo567 In Vitro Kit (RiboBio, Guangzhou, China), following the manufacturer’s protocol. The cells were incubated with 50 mM of EDU solution at 37 °C for 2 h, fixed with 4% paraformaldehyde, and then stained with the Apollo Dye reaction and Hoechst stain. Finally, the cells were observed and imaged using a fluorescence microscope, and the fluorescence results were analysed using ImageJ 1.8 (USA). The flow cytometry experiment was performed as follows: the digested cells were fixed with 75% of ethanol at −20 °C for 18 h, washed and added to the fluorescent dye (BD Cycletest™ Plus DNA Reagent Kit, San Jose, CA, USA), and then assayed using a flow cytometer (BD FACSVerseTM, Piscataway, NJ, USA), and the measurements were analysed using Modfit 3.1.
4
Quantifying Proliferation in Ovarian Cancer
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The CCK-8, colony formation, and 5-ethynyl-2’-deoxyuridine absorb assays were performed to detect OC cell proliferation in vitro. CCK-8 reagent (Yeasen, Shanghai, China) and BeyoClick™ EdU-488 kit were used to perform CCK-8 and 5-Ethynyl-2 absorb assays, respectively, according to the manufacturer’s instructions. For colony formation assay, 1000 OC cells were set in a 6-well plate and cultured at 37 °C for 14 days. The medium was then removed, and 4% paraformaldehyde was used to fix the cell colony. Finally, the cell colonies were stained with crystal violet, and colony number in each well was calculated.
5
Colon Cancer Cell Viability Assay
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The pretreated colon cancer cells were planted into a 96-well plate (1 × 103 cells/well). CCK-8 reagent (Yeasen Bio, shanghai, China) was co-incubated with the cells after 24, 48, 72, 96, and 120 h, respectively, according to the manufacturer’s instruction. OD450 values were determined via a microplate reader.
6
CCK-8 Cell Viability Assay
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RSC96 cells were seeded onto 96-well plates at an appropriate density (appropriately 30%). After 48 h of transfection, 10 μL of CCK-8 reagent (40203ES60, YEASEN Biotech, Shanghai, China) was injected into each well, followed by the inoculation of cells for 2 h at 37 °C with 5% CO2. To measure cell viability, the absorbance at 450 nm was detected using a microplate reader (Synergy 2, BioTek, USA).
7
Cell Viability Assay with CCK-8
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HEI-OC1 cells were collected, washed after the treatment, and plated on a 96-well plate at a density of 10,000 cells/well. The CCK-8 reagent (40203ES76; YEASEN, China; 10 μL), which can undergo a reduction reaction with succinate dehydrogenase in the mitochondria of living cells, was added to HEI-OC1 cells for 4 h of incubation (conventional culture conditions). The 96-well plate was placed in a microplate reader to analyze the absorbance at 450 nm.
8
Cell Viability Assay with StemRegenin 1 and CH-223191
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Cells were grown in the presence of different doses of StemRegenin 1 (0, 62.5, 125, 250, 500, 1,000, 2000, 4,000 nM) or CH-223191 (0, 7.8, 15.6, 31.3, 62.5, 125, 250, 500 nM) for 48 h. 10 μL of CCK8 reagent (Yeasen, Shanghai, China) was added, incubate for 2 h, and the absorbance measured at 450 nm. Using GraphPad Prism v. 8.0 (GraphPad Software, CA, United States), the half-maximal inhibitory concentration (IC50) was determined based on the relative survival curve.
9
Cell Viability Assay with BLU9931 and Palbociclib
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Cells (2,500 in 100 μl medium) were seeded into each well (N = 5) of a 96-well plate, and changed to medium with different concentrations of BLU9931 or palbociclib on the second day. After 72 h, 10 μl CCK8 reagent (YEASEN, #40203) was added and the optical density was measured at OD450 nm with a microplate reader (BioTek) after 1–4 h incubation.
10
CCK-8 Assay for Cell Proliferation
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The CCK-8 assay was performed to assess cell proliferation. Transfected cells were seeded into 96-well plates at 2 × 103 cells/well. 10 μL of CCK-8 reagent (40203ES80, YEASEN, China) was added to the wells after 24 h, 48 h and 72 h, and incubated for 4 hours at 37°C. Absorbance at 450 nm was measured using a microplate spectrophotometer (Bio-Tek, USA).
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