The MeT-5A cells were seeded into 96-well plates and transfected with NC, mimic, or inhibitor for 48 h. Each group of cells were then incubated with 10 µL CCK-8 reagents (Biosharp, Shanghai, China) for 2 h at 37 ℃. The viability of cells was measured by absorbance values at 450 nm.
Cck 8 reagent
Manufactured by Biosharp
Sourced in China, United States
The CCK-8 reagent is a colorimetric assay used to measure the viability and proliferation of cells. It utilizes a tetrazolium salt that is reduced by living cells, resulting in a change in color that can be quantified using a spectrophotometer.
Automatically generated - may contain errors
Lab products found in correlation
Microplate reader, by Agilent Technologies (4 mentions)
Spectramax m2, by Molecular Devices (2 mentions)
Microplate reader, by PerkinElmer (2 mentions)
Microplate reader, by Tecan (2 mentions)
Edu detection kit, by RiboBio (1 mentions)
Tl4 photomicroscope, by Olympus (1 mentions)
Compusyn software, by ComboSyn (1 mentions)
Penicillin streptomycin, by Sangon (1 mentions)
24 well companion plate, by Corning (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Phanta max super fidelity dna polymerase, by Vazyme (1 mentions)
Reverse transcription kit, by Toyobo (1 mentions)
Rapamycin, by Selleck Chemicals (1 mentions)
Het 1a, by American Type Culture Collection (1 mentions)
Bic 1, by American Type Culture Collection (1 mentions)
Spark multimode microplate reader, by Tecan (1 mentions)
Multiscan fc, by Thermo Fisher Scientific (1 mentions)
M200 pro, by Tecan (1 mentions)
A003 4 1, by Nanjing Jiancheng (1 mentions)
A020 2 2, by Nanjing Jiancheng (1 mentions)
48 protocols using cck 8 reagent
1
Cell Viability Assay with miRNA
2
Measuring Cell Proliferation via CCK-8 and EdU
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CCK-8 (Cell Counting Kit-8) assay was carried out as follows: 5,000 cells were seeded in each well of the 96-well plates and cultured overnight at 37°C. After treating with myricetin for 24 h, the cells were incubated with CCK-8 reagents (Biosharp, Beijing, China) for 1 h at 37°C. The light absorbance at 450 nm was measured by a microplate reader (SpectraMax M2, Molecular Devices, Shanghai, China).
The EdU (5-Ethynyl-2′-Deoxyuridine) proliferation assay was determined by using EdU detection kits (#C10310-1, Ribobio, Guangzhou, China) according to the manufacturer’s instruction. 5,000 cells were seeded in 96-well plates per well and cultured overnight. The cells were then treated with myricetin for 24 h. Then, the cells were stained with 50 μM EdU medium for 2 h, immobilized with 4% polyoxymethylene for 30 min, treated with 2 mg/ml glycine for 5 min, and permeated by 0.5% TritonX-100 for 10 min. After that, the cells were mixed with Apollo 567 for 30 min at room temperature in the dark and permeated again by 0.5% TritonX-100 for 20 min. Last, cells were maintained in Hoechst 33,342 for 30 min at room temperature in the dark. Cells were detected by the fluorescence microscope (Olympus TL4 photomicroscope, Japan). Proliferative cell nuclei were stained red fluorescence, and all cell nuclei showed blue fluorescence. The cell proliferation rate was analyzed by ImageJ.
The EdU (5-Ethynyl-2′-Deoxyuridine) proliferation assay was determined by using EdU detection kits (#C10310-1, Ribobio, Guangzhou, China) according to the manufacturer’s instruction. 5,000 cells were seeded in 96-well plates per well and cultured overnight. The cells were then treated with myricetin for 24 h. Then, the cells were stained with 50 μM EdU medium for 2 h, immobilized with 4% polyoxymethylene for 30 min, treated with 2 mg/ml glycine for 5 min, and permeated by 0.5% TritonX-100 for 10 min. After that, the cells were mixed with Apollo 567 for 30 min at room temperature in the dark and permeated again by 0.5% TritonX-100 for 20 min. Last, cells were maintained in Hoechst 33,342 for 30 min at room temperature in the dark. Cells were detected by the fluorescence microscope (Olympus TL4 photomicroscope, Japan). Proliferative cell nuclei were stained red fluorescence, and all cell nuclei showed blue fluorescence. The cell proliferation rate was analyzed by ImageJ.
3
Cell Proliferation Assays: CCK-8 and Colony Formation
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Cell proliferation was assessed by CCK-8 and colony formation assays. For the CCK-8 assay, stably transfected cell lines seeded into 96-well plates at 1 × 103 cells per well at a concentration of 100 μL, with five replicates for each condition. After incubation for an additional 0, 24, 48 or 72 h., 10μL CCK-8 reagents (Biosharp, China) were added to each well, and the cells were cultured at 37 °C for 2 h. The OD value was then measured at 450 nm using spectrophotometer. For colony formation assays, stably transfected cell lines seeded (1 × 103 cells/well) into 6-well plate, After1-2 weeks of culture, the colonies were fixed methanol, stained with crystal violet and analyzed using ImageJ software.
4
Microglia-Mediated HT22 Cell Viability
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Primary microglia were seeded into a 96-well plate (1 × 104 cells/well), and LPS was added to the cells for 4 h and then stimulated with 10 μM nigericin for 45 min. HT22 cells were plated in 96-well plates at a density of 103 cells per well, cultured for 24 h, then replaced the media with microglia-CM, and cultured for another 12 h. Finally, 10 µL CCK-8 reagents (Biosharp, China) were added per well. The absorbance at 450 nm was measured to calculate the cell viability using a microplate reader.
5
Cell Adhesion Assay on Fibronectin
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A 96‐well plate was coated with 50 μL FN (10 μg/mL) overnight at 4°C. Coated wells were then blocked with 0.2% BSA in PBS for 1 h at 37°C. Cells were seeded into 96‐well plates at 3 × 103 cells per well at a concentration of 100 μL and the adhesive ability of cells was analyzed at time points of 0, 30, 60, and 120 min. Nonadherent cells were removed by gentle washing with PBS. The remaining adherent cells were quantified using the CCK‐8 reagents (Biosharp).
6
Synergistic Effects of NaB and 5-FU
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Cell viability was assessed through the CCK-8 assay. Cells, including HCT-116, SW-480, DLD-1, and NCM-460, were seeded in 96-well plates with a density of 5 × 10 3 cells per well and cultured for 24 h. Upon reaching 70%-80% confluence, treatments were administered, involving various concentrations of NaB (0, 1, 2, 4, 8, 16, 32, and 64 mM), 5-FU (0, 1, 2, 4, 8, 16, 32, and 64 µg/mL), or a combination of 5-FU and NaB for an additional 24 or 48 h. Following this, 10 μL of CCK-8 reagent (Biosharp) was introduced to each well, and the incubation continued at 37 °C for 2 h. The ultimate absorbance at 450 nm was measured using a microplate reader (Perkin Elmer, Waltham, MA, USA). CompuSyn software (CompuSyn Inc., Paramus, NJ, United States) was engaged to calculate combination index (CI) values for the two drugs, and CI values below 1 signify synergistic effects.
7
Cell Viability Assay with SIN
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Cells were inoculated into 96-well plates at 5,000 cells per well and incubated for 24 h at 37°C. Cells were then treated with SIN for 24 h. CCK-8 reagent (Biosharp, Beijing, China) was added to each well and reacted with cells for 1 h at 37°C following the manufacturer’s instructions. Light absorbance at 450 nm was measured by a microplate reader (SpectraMax M2, Molecular Devices, Shanghai, China).
8
Cell Proliferation Assessment via CCK-8
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CCK-8 assay was performed to evaluate the proliferative ability of cells. Briefly, 1 × 103 cells in different treatment conditions were seeded into each well of a 96-well plate. A volume of 10 μL CCK-8 reagent (Biosharp, Shanghai, China) was added to each well and incubated for 1 h. After a brief shaking of the plate, the absorbance at 450 nm was assessed using a Varioskan LUX machine (ThermoFisher, Waltham, MA, USA).
9
Cell Viability Assay with ART
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The cells were inoculated in 96-well cell culture plates in appropriate quantities. After 24 h of incubation with the different concentrations of ART (Selleck, China), 10 μL CCK8 reagent (Biosharp, China) was added to each well. The plate was incubated for 4 h at 37°C in a CO2 incubator. The optical density (OD) of each well at 450 nm was measured by a Microplate Reader (BIOTEK, USA).
10
Cell Viability Assay for Transfected Cell Lines
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After transfection for 24 h, HepG2 and SNU‐387O2 cell lines were separately transferred to a 96‐well plate at a density of 5000 cells per well, with PBS covering the perimeter to prevent evaporation. Upon observing cell adhesion 24 h later, CCK‐8 reagent (Biosharp, China) was mixed with DMEM complete medium according to the manufacturer's instructions, and 200 μL of the mixture was added to each well to avoid bubble formation. The 96‐well plate was wrapped in aluminium foil to shield it from light and placed in an incubator. After a 3‐h incubation period, absorbance at 450 nm for each well was measured using a microplate reader. The procedure was repeated at 24, 48, 72 and 96‐h time points. Three replicate wells were set up for each group. This analysis aimed to ascertain whether there were statistically significant differences among the groups.
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