Antigen-specific splenocyte proliferation was evaluated by CCK-8 kits (Dojindo, Japan) according to the manufacturer's instructions. Splenocytes (5.0 × 106 cells per mL), stimulated with HBsAg (5 μg mL−1) or not, were seeded in triplicate (40 μL per well) in a 96-well plate and incubated at 37 °C in a humid atmosphere with 5% CO2. After 48 h, 20 μL of CCK-8 solution (Dojindo, Japan) was added into each well, and the plates were incubated for an additional 4 h. The cell proliferation was determined by CCK-8 kit, and the reagent of WST-8 (2-(2,4-nitrophenylmethoxy)-3-(4-nitrophenyl)-5-(2,4-disulfonic acid benzene)-2H-tetrazolium monosodium salt) could be reduced into water-soluble yellow formazan product (formazan dye) by dehydrogenase in cells under electronic carrier of 1-methoxy-5-methylphenazine dimethyl sulfate (1-methoxy PMS), which was in direct proportion to the number of living cells. Formazan product absorbed at 450 nm, and the absorbance at 450 nm (with 620 nm as reference) was measured by an Infinite M200 microplate spectrophotometer. The results were expressed as the proliferation index (PI) calculated based on the formula: PI = OD (450 nm) for stimulated cultures/OD (450 nm) for non-stimulated cultures.
Cck 8 kit
Manufactured by Dojindo Laboratories
Sourced in Japan, China, United States, Finland
The CCK-8 kit is a colorimetric assay for the determination of cell viability and cytotoxicity. It utilizes the highly water-soluble tetrazolium salt, WST-8, which is reduced by dehydrogenases in living cells to produce a colored formazan dye. The amount of the formazan dye generated is directly proportional to the number of living cells.
Automatically generated - may contain errors
Lab products found in correlation
Microplate reader, by Thermo Fisher Scientific (60 mentions)
Microplate reader, by Bio-Rad (59 mentions)
Microplate reader, by Agilent Technologies (42 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (34 mentions)
Microplate reader, by Molecular Devices (19 mentions)
Microplate reader, by Tecan (17 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (12 mentions)
96 well plate, by Corning (12 mentions)
Elx800, by Agilent Technologies (11 mentions)
Multiskan fc, by Thermo Fisher Scientific (10 mentions)
Crystal violet, by Beyotime (9 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (7 mentions)
Crystal violet, by Merck Group (7 mentions)
Streptomycin, by Thermo Fisher Scientific (7 mentions)
Multiskan go, by Thermo Fisher Scientific (6 mentions)
Trizol reagent, by Thermo Fisher Scientific (6 mentions)
Spectramax m5, by Molecular Devices (6 mentions)
Microplate spectrophotometer, by Bio-Rad (6 mentions)
Microplate reader, by Promega (6 mentions)
Trizol, by Thermo Fisher Scientific (6 mentions)
1 386 protocols using cck 8 kit
1
Splenocyte Proliferation Assay with CCK-8
2
Evaluating Ferroptosis Inducers and FAAH Inhibitors
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786-O cells were seeded in 96-well plates for each cell line at 4000 cells per well, and a cell viability assay was conducted with a CCK-8 kit (Dojindo Laboratories, Japan) after combinational treatment with FAAH inhibitors and ferroptosis inducers for 72 h according to the manufacturer’s instructions. For the drug synergy assay, the synergic effect of drug pairs was defined quantitatively with the Chou–Talalay equation [46 (link)], where a combination index < 1 indicates synergism and a combination index > 1 indicates antagonism. 786-O and Caki-1 cells were treated with different concentrations of two single drugs and combinational treatment, respectively, for 72 h, and cell viability was validated with a CCK-8 kit (Dojindo Laboratories) according to the manufacturer’s instructions.
3
Scaffold-Mediated Cell Proliferation
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Briefly, mBMSCs (ATCC Inc., USA) and HUVECs (Sciencell Inc., USA) were seeded at the density of 105 per scaffold in 48-well plates for 24 h using complete medium (high glucose Dulbecco’s modified Eagle’s medium (H-DMEM) with 10% fetal bovine serum). Then, the original mediums in the plates were replaced with new mediums. The influences of the scaffolds on the proliferation of mBMSCs and HUVECs were then assessed by using CCK-8 kit (Dojindo Molecular Technologies Inc., Japan) at day 1, 3, and 5. The cell proliferation was detected by CCK-8 kit (Dojindo Molecular Technologies Inc., Japan) after seeding cells onto the scaffold for 1, 3, and 5 d.
4
Cell Proliferation Assay for HCC4006 and NCI-H1703
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Proliferation of HCC4006 and NCI-H1703 cells after transfections were detected using CCK-8 kits (Dojindo, Japan) [18 (link)]. After washed with PBS, 5000 cells were transferred to each well of a 96-well plate and cultured at 37°C. Cell numbers were reflected by measuring OD values of medium at 450 nm at 2 h after the addition of 10% CCK-8. Measurements were performed every 24 h for a total of 96 h.
5
CoNPs Cytotoxicity and Antioxidant Assays
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CoNPs (< 50 nm, #7440-48-4), alpha-lipoic acid (#1077-28-7), and dichlorofluorescin diacetate (DCFH-DA) (#4091-99-0) were purchased from Sigma-Aldrich (St. Louis, MO). Dulbecco’s modified Eagle's medium (DMEM), fetal bovine serum (FBS), penicillin/streptomycin, and trypsin–EDTA were procured from Gibco (Life Technologies, Paisley, UK). CCK-8 kits were obtained from Dojindo (Kumamoto, Japan). Annexin V-FITC (#AP101-100-AVF), Calcein AM/PI (#C2013FT& ST511) was purchased from Multi Sciences (Hanzhou, China). DMSO, GSH, and GSSG Assay Kit (#S0053) were procured from Beyotime (Shanghai, China). Iron Assay Kit (#ab83366), Anti-GPx4 antibody (#ab125066), Goat anti-rabbit IgG H&L (HRP) (ab205718), Goat anti-mouse IgG H&L (HRP) (ab205719), and β-actin antibody were obtained from Abcam Technology (Cambridge, UK).
6
Cell Viability Assay using CCK-8
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Cell viability was detected using CCK-8 kits (DOJINDO, Japan) according to the manufacturer’s protocols. Briefly, cells were plated into 96-well plate. At the indicated time after treatments, 10 μl of CCK-8 solution was added to the wells. Cells were then incubated at 37°C for 1 h. Finally, a microplate reader (Labsystems Dragon, Finland) was used to measure the absorbance at 450 nm.
7
Evaluating Chemoresistance in Tumor Cell Lines
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CCK-8 kits (CK04, Dojindo, Kumamoto, Japan) were utilized for evaluating the proliferative potential of MUM-2B and MUM-2B/CDDP cells. MUM-2B and MUM-2B/CDDP cells in the logarithmic growth phase were seeded in a 96-well plate (1 × 104 cells per well) and cultured for 1, 2, 3, 4, and 5 days. During this period, 10 μL CCK-8 reagent was loaded at the same time every day, and incubation was made at 37 °C for 3 h. The OD value of each well at 450 nm was measured using a microplate reader. The values were proportional to the number of proliferated cells in the medium, and the cell growth curve was plotted.
MUM-2B and MUM-2B/CDDP cells in logarithmic growth phase were seeded in a 96-well plate (1 × 104 cells per well) for 24 h. After 24 h, cells were exposed to different concentrations of CDDP. After 48 h, 10 μL CCK-8 reagent was loaded for another 3 h of incubation at 37 °C. The OD value of each well at 450 nm wavelength was recorded on a Microplate reader and finally presented in the cell growth curve. IC50 refers to the concentration of cisplatin inhibiting the cell viability by 50%. A higher IC50 value indicates higher resistance to chemotherapy [26 (link)].
MUM-2B and MUM-2B/CDDP cells in logarithmic growth phase were seeded in a 96-well plate (1 × 104 cells per well) for 24 h. After 24 h, cells were exposed to different concentrations of CDDP. After 48 h, 10 μL CCK-8 reagent was loaded for another 3 h of incubation at 37 °C. The OD value of each well at 450 nm wavelength was recorded on a Microplate reader and finally presented in the cell growth curve. IC50 refers to the concentration of cisplatin inhibiting the cell viability by 50%. A higher IC50 value indicates higher resistance to chemotherapy [26 (link)].
8
Cytotoxicity Evaluation of Peptides
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HL‐60 and HEK 293 were used in CCK‐8 assays for the evaluation of cytotoxic effects of peptides. Cells were cultured at 37°C in 5% CO2: 95% air incubator in Minimum Essential Medium (MEM) supplemented with 10% (v/v) fetal bovine serum (FBS). Cells were seeded at a density of 7 × 103 cells per well in 96‐well plates and incubated for 24 h. Media were then replaced with 200 μl of media containing various concentrations of peptides. The cells were incubated for 72 h and cytotoxicity was assessed with CCK‐8 Kits (Dojindo Molecular Technologies, Tokyo, Japan). Absorbance was detected at 450 nm with the TECAN Infinite M200 microplate reader (Tecan, Durham, USA).
9
Cell Viability Assay of Liver Cancer Cells
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Hep3B as well as HUH-7 cells were planted onto 96-well plates (3,000 cells/well). In line with the protocols of CCK-8 kits (Dojindo, Japan), 10 μL CCK-8 solution that was diluted by 100 μL DMEM replaced the previous DMEM at diverse hours (24, 48, 72, and 96 h). After being cultured protecting from light at 37°C lasting an extra two hours, viable cells were determined through absorbance at 490 nm wavelength.
10
Resveratrol and SRT1720 Activate Mitochondrial Biogenesis
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Cells were treated with 1 mM H2O2 (Sigma-Aldrich, USA) for 2 h. To activate mitochondrial biogenesis, cells were preincubated with 5 μM resveratrol (Sigma-Aldrich) or 0.5 μM SRT1720 (Selleck, USA) for 24 h. Cell viability was detected using CCK-8 kits (DOJINDO, Japan) according to the manufacturer's protocols. Briefly, cells were plated into a 96-well plate. At the indicated time after treatments, 10 μl CCK-8 solution was added into the wells. Cells were then incubated at 37°C in an incubator for 1 h. Then, a microplate reader (Labsystems Dragon, Finland) was used to measure the absorbance at 450 nm.
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