A Cell Counting Kit-8 (CCK-8) kit (Roche Diagnostics) was used for the cell viability assay according to the manufacturers instructions. Cells (1×103 cells/well) were transiently transfected and seeded in 96-well plates overnight at 37°C, and then treated with stimulants for 3 days. CCK-8 (10 µl) was then added and the cells were incubated at 37°C for a further 4 h. Colorimetric absorbance was measured at 450 nm using an ELISA microplate reader, with six replicates per experimental sample.
Cell counting kit 8 (cck8)
Manufactured by Roche
Sourced in Switzerland, United States, Germany
The Cell Counting Kit-8 is a colorimetric assay for the determination of the number of viable cells in cell proliferation, cytotoxicity, and survival studies. The kit uses WST-8, a water-soluble tetrazolium salt, to measure the number of living cells. The assay is based on the reduction of WST-8 to a water-soluble formazan dye, which is directly proportional to the number of viable cells. The resulting color change can be measured using a spectrophotometer.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
96 well plate, by Corning (1 mentions)
Biotek gen5 system, by Bioteck (1 mentions)
6000 system, by Rayto (1 mentions)
Dapi 4 6 diamidino 2 phenylindole, by Beyotime (1 mentions)
Sybr green realtime pcr master mix, by Toyobo (1 mentions)
In situ cell death detection kit, by Roche (1 mentions)
Heat inactivated fetal bovine serum, by Biowest (1 mentions)
Rpmi 1640 medium with l glutamine, by Merck Group (1 mentions)
Low melting point agarose lmp, by Merck Group (1 mentions)
Hydrogen peroxide, by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Matrigel, by BD (1 mentions)
14 protocols using cell counting kit 8 (cck8)
1
Cell Viability Assay using CCK-8
2
Cell Proliferation and Cisplatin IC50
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Cell proliferation and cisplatin IC50 were determined using the Cell Counting Kit-8 (CCK-8, Roche, Basel, Switzerland) and following the manufacturer’s instructions. The absorbance was detected spectrophotometrically at 450 nm.
3
Cell Proliferation and Colony Formation Assay
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Cell proliferation was measured by Cell Counting Kit-8 (CCK-8, Roche, Basel, Switzerland) following the manufacturer’s instructions. Cells (2 × 103/well) were seeded into 96-well plates (Corning, NY, USA) and incubated for 24 h, 48 h, and 72 h, respectively. 10 μL of CCK-8 regents were added into cells and incubated for 2 h at 37 °C. The absorbance was measured at 450 nm on the spectrophotometer.
Colony formation assay was conducted for 14 days, and the colonies were fixed in 70% ethanol for 10 min and then stained with 1% crystal violet solution for another 10 min at room temperature. Images were captured with a camera (Nikon, Japan).
Colony formation assay was conducted for 14 days, and the colonies were fixed in 70% ethanol for 10 min and then stained with 1% crystal violet solution for another 10 min at room temperature. Images were captured with a camera (Nikon, Japan).
4
Cell Viability Assay on 96-well Plates
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Cells were plated on 96-well plates. As the indicated time points, the viability of cells was determined by Cell Counting Kit 8 (Roche) and measured at OD 450 nm with the BioTek Gen5 system (BioTeck, USA). The experiments were repeated three times with the representative experiment shown here.
5
Cell Viability Assessment using CCK-8 Assay
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Cell Counting Kit-8 (Roche Biochemicals, Germany) was used for cell viability measurement. 100 μl cell suspensions (2 × 104 cells per ml) with 10% FBS medium was seeded to a 96-well plate and incubated for 5 days at 37°C. Then, 10 μl CCK-8 reagent was added to each well at the indicated time and incubated for 4 h at 37°C. Absorbance was measured at 450 nm by a Rayto-6000 system (Rayto, China).
6
Assessing Proliferation of RCC Cells
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Cell Counting Kit-8 (CCK-8; Roche, Basel, Switzerland) was used to assess the proliferation of RCC cells. After seeding 1×104 transfected cells in 96-well plates, they were cultured for 24, 48 or 72 hours. 10μl of CCK-8 was then added to the cells and cultured for 2 hours. The absorbance of the plate was read at 450nm using a microplate reader.
7
Docetaxel-resistant Breast Cancer Cell Lines
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Human breast cancer cell lines MDA-MB-231, docetaxel (TXT)-resistant SKBR3, TXT-resistant BT474, TXT-resistant MCF7, and TXT-resistant MDA-MB-231 were obtained from Dr. Wang HH. All of these cell lines were cultured according to the ATCC instructions. siRNA and Lipofectamine™ 2000 were purchased from Thermo Scientific Dharmacon®. SYBR Green Real-time PCR Master Mix was purchased from Toyobo (Osaka, Japan). RevertAid™ First Strand cDNA Synthesis kit was obtained from MBI (Fermentas, Hanover, MD, USA). Docetaxel (Aventis Pharmaceuticals, Bridgewater, NJ, USA) was stored at a concentration of 10 mg/mL (12.6 mM) in 13% w/w ethanol at 4 °C and diluted in medium before use. Cell Counting Kit-8 was obtained from Roche. TUNEL assay was performed by use of In Situ Cell Death Detection Kit (Fluorescein, Roche, Switzerland). DAPI (4,6-diamidino-2-phenylindole) was purchased from Beyotime (Shanghai, China). Hoechst 33342 was purchased from Sigma-Aldrich China (Shanghai, China).
8
Cell Proliferation Assay using CCK8
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Cell Counting Kit-8 (CCK8; Roche, Switzerland) was used to determine cell proliferation. 10 μl of CCK8 solution was added into each well of 96-well plates containing 5 × 103 cell. After 2 h, a microplate reader (Thermo scientific, USA) was used to measure absorbance at 450 nm.
9
Cytotoxicity Evaluation of Cisplatin
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Cell Counting Kit-8 (CCK-8), cisplatin (cisPt), Cell Proliferation ELISA - BrdU colorimetric Kit (Roche), DAPI (4′,6-diamidino-2-phenylindole dihydrochloride), 2′,7′-dichlorofluorescin diacetate (H2DCF-DA), penicillin-streptomycin solution, normal melting-point agarose (NMP), RPMI-1640 medium with L-glutamine, Triton™ X-100, hydrogen peroxide, and low melting-point agarose (LMP) were purchased from Sigma-Aldrich Chemicals (St. Louis, MO, USA). Dulbecco’s Phosphate Buffered Saline without calcium and magnesium (DPBS), Hanks BSS (HBSS) without phenol red with calcium and magnesium were purchased from Biological Industries (Cromwell, CT, USA), and heat-inactivated fetal bovine serum (FBS) from Biowest (Nuaillé, France). All other analytical grade and high-quality chemicals were obtained from local commercial suppliers, such as Chempur (Piekary Slaskie, Poland) or POCH S.A. (Gliwice, Poland).
10
Cell Viability Assay for miR-29 Transfected NSPCs
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Cell viability was evaluated as previously described (12 (link)) using a Cell Counting Kit-8 (Roche Applied Science) according to the manufacturer's protocols. For the miR-29 group, NSPCs were grown in 24-well plates, the other groups were grown in 96-well plates at 5,000 cells/well for 24 h before transfection. Cell viability was detected at the end of each treatment (6, 12, 24, 48 and 72 h following transfection) using an Epoch multi-microplate spectrophotometer (Bio-Tek Instruments, Inc.).
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