The CCK8 (Absin, Shanghai, China) was utilized to assess cellular viability. In short, cells were seeded onto 96-well plates at a density of 5,000 cells per well. Once the cells reached 60% confluence, they were treated with either sorafenib (10 μM) or dimethyl sulfoxide (DMSO) for a duration of 24 h. Following this, 10 μL of CCK-8 solution was introduced into each well and incubated for 2 h. Subsequently, the absorbance at 450 nm was determined using a microplate reader.
Cell counting kit 8 (cck8)
Manufactured by Absin
Sourced in China
The CCK8 is a colorimetric assay kit used to measure cell viability and cytotoxicity. It utilizes a tetrazolium compound that is reduced by metabolically active cells, producing a colored formazan dye that can be quantified spectrophotometrically.
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Lab products found in correlation
Spectramax m5, by Molecular Devices (2 mentions)
96 well microplate, by Corning (1 mentions)
Annexin 5 fitc apoptosis detection kit, by Beyotime (1 mentions)
Live dead cell imaging kit, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trypsin edta 1x, by Thermo Fisher Scientific (1 mentions)
N n dimethylformamide (dmf), by Maclin Biochemical Technology (1 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions)
Rsc 96, by National Collection of Authenticated Cell Cultures (1 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Cell light edu apollo488 in vitro kit, by RiboBio (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Chitosan, by Maclin Biochemical Technology (1 mentions)
Milli q apparatus, by Merck Group (1 mentions)
Carboxyfluorescein diacetate succinimidyl ester, by MedChemExpress (1 mentions)
Rpmi 1640 media, by Sartorius (1 mentions)
Fitc phalloidin and dapi, by Solarbio (1 mentions)
13 protocols using cell counting kit 8 (cck8)
1
CCK-8 Assay for Cell Viability
2
Cell Viability Assay Using 5D3PC
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Cells seeded at a density of 5 × 103 cells/well in 96-well were incubated with 5D3PC for 24 h. Then the culture medium was replaced with the CCK-8 (Absin, Shanghai, China) solution (10 µL), and the cells were further cultured for 2 h at 37 °C. Plates were read at 450 nm using a microplate reader Spectra Max M5 (Molecular Devices company, San Diego, CA, USA). An equal volume of cells without the treatment was used as a blank control. All the experiments were repeated three times.
3
Methyl Vanillate Cytotoxicity Evaluation
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CCK8 (Absin) was used to determine cell viability. The cells were plated in 96-well microplates (Corning) with a seeding density of 5×103 per well and 100 µL of the medium. Cells were treated with methyl vanillate at different concentrations (100, 200, 400, 800, and 1,600 µmol/L) for 24 hours. CCK8 reagent (10 µL) was added to each well and then cultured for 2 hours. All experiments were performed 3 times. The absorbance was measured at 450 nm using a microplate reader with a blank well (SPECTROstar). The proliferation of cells was reflected in the absorbance.
4
Measuring Cell Viability in SiHa Cells
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A 96-well plate was used for the stable SiHa cells growing at a density of 3 × 103 cells per well, and the plate was removed after pre-culture for 24 h. Then, 10 μL CCK-8 (Absin, Shanghai, China) was put into each well under dark conditions and 1 h in an incubator. We monitored the OD450 value at five time-points (0, 24, 48, 72, and 96 h) using a microplate reader.
5
PCL-MLT-RGO Nanoparticle Cytotoxicity Assay
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The PCL (30 kDa) was purchased from HENGQIU (Suzhou, China). The MLT was purchased from Sigma-Aldrich (Shanghai, China), and the RGO nanoparticles were purchased from XFNANO (Nanjing, China). DCM was purchased from Aladdin (Shanghai, China), and DMF was purchased from Macklin (Shanghai, China). The RSC 96 were provided by the National Collection of Authenticated Cell Cultures (Shanghai, China). The Fetal Bovine Serum (FBS), penicillin/streptomycin solution, 0.25% Trypsin-EDTA (1x), and Dulbecco's Modified Eagle's Medium (DMEM) with 4.5 g/L d -Glucose were purchased from Gibco (USA). The LIVE/DEAD Cell Imaging Kit was purchased from Thermo Fisher Scientific (USA), the Annexin V-FITC Apoptosis Detection Kit was purchased from Beyotime (China), and the JC-1 Mitochondrial Membrane Potential Assay Kit was purchased from MCE (Shanghai, China). The CCK8 was purchased from Absin (Shanghai, China), and all the antibodies were purchased from Abcam (USA). The 3–0 nylon sutures, 6–0 nylon sutures, and sterile scalpel blades were purchased from Jinhuan Medical (Shanghai, China).
6
Evaluating Proliferation of A2780 and OVCAR8 Cells
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The proliferation of A2780 and OVCAR8 cells was evaluated using CCK8(Absin, Shanghai, China) according to the instructions. Briefly, cells were resuspended and counted, then seeded at 1 × 103/well in a 96-well plate. The medium was aspirated after 12 h, then 10 μL of CCK8 solution was added to the new medium. After incubating for 2 h at 37 °C in a 5% CO2 atmosphere, the absorbance was measured at a wavelength of 450 nm, and the time was recorded as 0 h. Readings were repeated on days 1, 2 3 and 4.
7
Evaluation of B16 Cell Viability
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The viability of the B16 cells was evaluated using a CCK-8 assay as previously described [40 (link)]. Briefly, the B16 cells were seeded at a density of 5 × 103 cells per well in 96-well and incubated with CC7 at a concentration of 1–50 µM for 24 h. Then, the culture medium was replaced with the CCK-8 (Absin, Shanghai, China) solution (10 µL). Then, the cells were further incubated for 2 h at 37 °C, and the absorbance was read at 450 nm using a microplate reader Spectra Max M5 (Molecular Devices Company, San Diego, CA, USA). An equal volume of cells without treatment was used as a blank control. All the experiments were repeated three times.
8
Cell Proliferation and Viability Assays
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Cell counting Kit-8 (absin, Shanghai, China) was used to detect cell proliferation. Proper concentration cells were suspended in 96 wells standard culturing plate. After 8 h of culturing, 10% CCK solution was added to the well, followed by incubating for 40 min. Then the OD 450 nm was collected by a microplate reader.
EdU incorporation was assayed using a Cell-Light EdU Apollo488 In Vitro Kit (RiboBio, Guangzhou, China) according to the manufacturer’s instructions. Briefly, Treated or untreated HUVECs were labeled with EdU for 2 h, fixed with 4% paraformaldehyde for 20 min, neutralized with 2 mg/ml glycine for 5 min, permeation with 0.5% TritonX-100 for 10min, stained with Apollo® buffer for 10min at room temperature. EdU incorporation was quantified by measuring the absorbance at 488 nm by a microplate reader (Agilent, Hangzhou, China).
EdU incorporation was assayed using a Cell-Light EdU Apollo488 In Vitro Kit (RiboBio, Guangzhou, China) according to the manufacturer’s instructions. Briefly, Treated or untreated HUVECs were labeled with EdU for 2 h, fixed with 4% paraformaldehyde for 20 min, neutralized with 2 mg/ml glycine for 5 min, permeation with 0.5% TritonX-100 for 10min, stained with Apollo® buffer for 10min at room temperature. EdU incorporation was quantified by measuring the absorbance at 488 nm by a microplate reader (Agilent, Hangzhou, China).
9
Cholesterol Depletion Effect on IPEC-J2 Cells
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The membrane cholesterol of IPEC-J2 cells was deleted by treating with different concentrations of MβCD (Sigma, St. Louis, MO, USA). The cell viability of IPEC-J2 cells after treating with MβCD was quantitated using a Cell Counting Kit-8 (Absin, Shanghai, China), as previously described [40 (link)]. Briefly, 100 μL of DMEM medium containing different concentrations of MβCD (0–20 mM) was added to each well of IPEC-J2 cells in the 96-well plate and incubated for 1 h at 37 °C. Then, 10 μL of CCK8 solution was added to each well. The absorbance values of the samples were read at 450 nm using a microplate reader. The percentage viability of cells was calculated with respect to that of the untreated control cells (set as 100%). Each concentration group had 12 replicate wells, and the experiment was performed three times.
10
Evaluating BCG-Induced Cell Viability
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Cell Counting Kit-8 was purchased from Absin (abs50003, Absin, Shanghai, China). THP-1 macrophages at the logarithmic growth stage were taken and treated with a cell density of 1 × 104·mL−1, then inoculated into 96 well culture plates. After culturing cells for 48 h and infecting them with BCG for 24 h, 10 μL of CCK-8 solution was added to each well. The absorbance (OD) of each well was measured at a wavelength of 450 nm using a fluorescence microplate reader, and data were recorded after 4 h. Cell viability (%) = (OD value of experimental group − OD value of blank group)/OD value of control group × 100%.
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