Cells were seeded into 96-well plates at a density of 1,000 cells per well. Then, 100 μl of cell culture medium was added to each well, and the cells were cultured in a 37°C, 5% carbon dioxide incubator. For detection, 10 μl of CCK-8 solution (Solarbio Science & Technology, China) was added to each well; the same volumes of cell culture medium and CCK-8 solution were added to blank control wells, but no cells were seeded. After incubation for 30 min, the absorbance was measured at 450 nm with a microplate reader.
Cck 8 solution
Manufactured by Solarbio
Sourced in China, United States
CCK-8 solution is a colorimetric assay that uses the tetrazolium salt WST-8 to measure cell viability and proliferation. It provides a simple and convenient method for determining the number of viable cells in cell proliferation and cytotoxicity assays.
Automatically generated - may contain errors
Lab products found in correlation
Microplate reader, by Thermo Fisher Scientific (18 mentions)
Microplate reader, by Bio-Rad (10 mentions)
Microplate reader, by Agilent Technologies (9 mentions)
Multiskan fc, by Thermo Fisher Scientific (6 mentions)
Microplate reader, by Allsheng (3 mentions)
Microplate reader, by Molecular Devices (2 mentions)
Multimode reader, by Agilent Technologies (2 mentions)
Hiscript 2 one step rt pcr kit, by Vazyme (2 mentions)
Mirna 1st strand cdna synthesis kit, by Vazyme (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
Aceq qpcr sybr green master mix, by Vazyme (2 mentions)
Synergy 2 multi mode microplate reader, by Agilent Technologies (2 mentions)
Microplate reader, by Bioteck (2 mentions)
Multiskan fc microplate reader, by Thermo Fisher Scientific (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Multiskan go, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Multiskan go microplate reader, by Thermo Fisher Scientific (1 mentions)
5 ethynyl 2 deoxyuridine (edu), by Abbkine (1 mentions)
140 protocols using cck 8 solution
1
Cell Viability Quantification with CCK-8
2
Cell Proliferation and Adhesion Assay
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To carry out the cell proliferation test, 1 × 106 HUCMSCs or NSCs were cocultured with 3D-CS-MExos and 3D-CS-HMExos for 1, 3, 5, and 7 days. The culture medium was replaced with CCK-8 solution (Solarbio) and incubated for 3 h. Then, the absorbance of the supernatant was measured at 450 nm with a microplate analyser (BioTech). The adhesion of NSCs to the scaffold was tested by the following method. Briefly, NSCs at a density of 1 × 105 per well were cocultured with 3D-CS-MExos and 3D-CS-HMExos for 3 days, fixed with 4% paraformaldehyde, and stained with F-actin and DAPI, and the morphology of the NSCs was detected by CLSM.
3
CCK-8 Cell Viability Assay
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The cells were seeded onto the 96-well plates, at a density of 1 × 105 cells/ml, in 100 μL culture medium each well. After being cultured in a 37°C, 5%CO2 incubator for 24 h, the cells were subjected to different treatments, respectively, for 7 days. Then, 10 μl CCK-8 solution (CA1210; Solarbio, Wuhan, Hubei, China) was added into each well to incubate the cells at 37°C for 4 h. The absorbance at 450 nm was added with the Multiskan GO microplate reader (Thermo, Waltham, Massachusetts, USA). Each group had 6 replicate wells, and the cell viabilities were calculated accordingly.
4
Cell Viability Assay using CCK-8
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The viability of the cells was observed by CCK-8 assay. 5 × 104 cells were seeded into the 96-well plates. After cell transfection, the viability of the cells at 0, 24, 48, and 72 hours were measured by CCK-8 assay. Briefly, 10 μL of CCK-8 solution (Solarbio Biotechnology Co., Ltd., Shanghai, China) was added into each well, and then, the cells were further cultured in the dark for 4 hours. The viability of the cells was observed by a microplate reader (Molecular devices, Shanghai, China) at 450 nm.
5
Cytotoxicity Evaluation of Cell-Penetrating Peptide
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The cytotoxic effect of CPP on lymphocytes was measured by Cell Counting Kit-8 (CCK-8) assay. Cells were treated with different doses of CPP (400, 200, 100, and 50 μg/mL), respectively. After incubation at 37°C for 48 h, 10 μL of the CCK-8 solution (CA1210; Solarbio, Beijing) was added to each well. The plates were reincubated at 37°C for 30 min, followed by measurement of absorbance values at 450 nm using a microplate reader (Bio-Rad, USA). The cell viability was calculated as follows: cell viability = OD450 nm values of cells with CPP treatment ÷ OD450 nm values of control cells without treatment.
6
Curcumin Protects Astrocytes from OGD/R
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The isolated astrocyte was seeded in a 96-well plate at a density of 8,000 cells per well and incubated at room temperature overnight. Curcumin (#C110685, Aladdin, China) was added at various concentrations (1 μM, 5 μM, 10 μM, 20 μM, and 40 μM) into the medium 30 min before OGD and at the time of reoxygenation. Approximately 24 h after oxygen-glucose deprivation/reoxygenation (OGD/R), 10 mL of CCK8 solution (#CA1210, Solarbio, China) was added and incubated in the dark for 1 h. The optical density values were measured at a wavelength of 450 nm.
7
Cell Viability and Proliferation Assays
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CCK-8 (CCK-8 solution: Solarbio, Beijing, China) was used to assess the viability of the cells. We seeded 2000 transfected cells in 96-well with 100 μl 10% FBS cell culture mediumand measured the optical density (OD) value after incubation at 37 °C for 3 hours. We detected the optical density value at 450 nm at the same time every day for four days. For colony formation assay, the transfected cells were seeded in 6-well plates with 300 cells per well supplemented with 2 mL 10% FBS cell culture medium and cultured for at least one week. The 6-well plates were fixed with 4% paraformaldehyde for 15 min and then stained with 0.1% crystal violet solution for 20 min. The EdU assay was also used to verify the proliferative ability of the cells. The transfected cells were incubated with EdU (Abbkine, China), and cell nuclei were stained with DAPI, and observed by fluorescence microscope. All experiments were repeated at least three times.
8
qRT-PCR and CCK-8 Assay Protocol
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After extracting RNA with Trizol (Invitrogen), RNA was reversely transcribed using HiScript II One Step RT-PCR Kit (Vazyme, Nanjing, China) or miRNA 1st Strand cDNA Synthesis Kit (Vazyme). Then, qRT-PCR was carried out using AceQ qPCR SYBR Green Master Mix (Vazyme). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and U6 were taken as internal controls. Primers were shown below: miR-198-F: 5'-GGTCCAGAGGGGAGAT-3', miR-198-R: 5'-GAATACCTCGGACCCTGC-3'; ABCB1-F: 5'-CCCATCATTGCAATAGCAGG-3', ABCB1-R: 5'-GTTCAAACTTCTGCTCCTGA-3'; GAPDH-F: 5'-TGCACCACCAACTGCTTAGC-3', GAPDH-R: 5'-GGCATGGACTGTGGTCATGAG-3'; U6-F: 5'-CTCGCTTCGGCAGCACA-3', U6-R: 5'-AACGCTTCACGAATTTGCGT-3'; 18sRNA-F: 5'-AAACGGCTACCACATCCA-3', 18sRNA-R: 5'-CACCAGACTTGCCCCTCCA-3'. The primers of circ_0002060 were purchased from GenePharma.
Cell Counting Kit-8 (CCK-8) assay Cells (3×10 3 ) were plated into 96-well plates and incubated with escalating doses of DOX for indicated time. Subsequently, cells were reacted with 10 μL CCK-8 solution (Solarbio) for 2 h. Next, a Multi-Mode Reader (BioTek, Burlington, VT, USA) was used to measure the optical density at 450 nm to assess cell viability. The half-maximal inhibitory concentration (IC50) of DOX is the concentration at which cell viability is reduced to 50%.
Cell Counting Kit-8 (CCK-8) assay Cells (3×10 3 ) were plated into 96-well plates and incubated with escalating doses of DOX for indicated time. Subsequently, cells were reacted with 10 μL CCK-8 solution (Solarbio) for 2 h. Next, a Multi-Mode Reader (BioTek, Burlington, VT, USA) was used to measure the optical density at 450 nm to assess cell viability. The half-maximal inhibitory concentration (IC50) of DOX is the concentration at which cell viability is reduced to 50%.
9
Cell Proliferation Assays for miRNA Transfection
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Cell proliferation was measured using the Cell Counting Kit-8 (CCK-8) and colony formation assays. After a 24-h transfection with miRNA, 5×103 transfected cells were seeded into each well in 96-well plates with 100 μL of medium. After 0, 24, 48, 72, and 96 h of incubation, 10 μL of the CCK-8 solution (Solarbio) was added to each well and incubated at 37°C for 1 h. Results were detected by a microplate reader with absorbance at 450 nm. For the plate colony assays, approximately 2×103 transfected cells were inoculated into each well in six-well plates. After two weeks of culture, the cells were fixed with 4% paraformaldehyde, stained using 0.1% crystal violet (Solarbio) for 10–30 min, and photographed after rinsing. Each experiment was performed three times.
10
Cell Proliferation Assay with CASC9 and PTEN Knockdown
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EJ and T24 cells were seeded onto 6-well plates and then transfected with CASC9 shRNA, PTEN shRNA or shRNA-NC as described above. At 48 h post-transfection, EJ and T24 cells were plated into a 96-well plate and incubated for 12 h in a 5% CO2 humidified incubator at 37°C. Following this, EJ and T24 cells were treated with CCK-8 solution (Solarbio) for 4 h and then incubated with formazan solution for 10 min on a shaker. The absorbance was determined at the wavelength of 450 nm.
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