At 24 h post-transfection, cells were harvested and seeded at 5×103 per well onto a 96-well plate. Cell proliferation was determined at day 1, 2, 3 and 4. According to the manufacturer's protocol, 10 µl Cell Counting Kit (CCK)-8 solution (Beyotime Institute of Biotechnology) was used for each coloring reaction. After a further incubation at 37°C for 2 h, the absorbance at 450 nm was quantified using a plate reader (Bio-Rad Laboratories, Inc.). Each experiment was performed in triplicate independently.
Cell counting kit 8 solution
Manufactured by Beyotime
Sourced in China, United States
The Cell Counting Kit-8 solution is a colorimetric assay for the determination of cell viability and cytotoxicity. The kit utilizes the water-soluble tetrazolium salt WST-8, which is reduced by dehydrogenases in living cells to produce a yellow-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 Bio-Rad (4 mentions)
Microplate reader, by Thermo Fisher Scientific (4 mentions)
Microplate reader, by Agilent Technologies (2 mentions)
Microplate reader, by Molecular Devices (2 mentions)
Microplate reader, by Tecan (1 mentions)
Rpmi 1640 medium, by Beyotime (1 mentions)
Infinite m200 pro, by Tecan (1 mentions)
Ldh assay kit, by Beyotime (1 mentions)
Thermo multiskan spectrum reader, by Thermo Fisher Scientific (1 mentions)
Ldh activity kit, by Beyotime (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Rpmi 1640 medium, by Sartorius (1 mentions)
Elx800, by Agilent Technologies (1 mentions)
Synergytm ht, by Agilent Technologies (1 mentions)
Crystal violet staining solution, by Beyotime (1 mentions)
25 protocols using cell counting kit 8 solution
1
Cell Proliferation Assay Using CCK-8
2
Cell Viability Assay with CCK-8
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Inoculated into 96-well plates at the density of 2 × 104 cells/well for incubation, BRL-3A cells were then added with 10 μl of Cell counting kit (CCK)-8 solution (Beyotime). After 2 h, the absorbance value of each hole was decided by a microplate reader (BioTek microplate reader) in the premise of λ = 450 nm.
3
Cell Proliferation Assay for NSCLC
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The NSCLC cells transfected with siRNA-GACAT1 or siRNA-control were cultured in 96 well plates, at a density of 1,000 cells per well and maintained at 37°C with 5% CO2. Then, 10 µl Cell Counting Kit (CCK)-8 solution (Beyotime Institute of Biotechnology) was added to the medium at daily intervals, including 1-4, and 5 days. The cells were incubated 37°C for an additional 4 h and the absorbance was detected at 450 nm using a microplate reader (Tecan Group, Ltd.). The experiment was performed with three replicates. Data was obtained from three independent experiments.
4
Endothelial cell viability in oxidative stress
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Human umbilical vein endothelial cells were purchased from the Cell Bank of the Type Culture Collection of the Chinese Academy of Sciences, Shanghai Institute of Cell Biology, Chinese Academy of Sciences, cultivated in endothelial cell medium. and divided into control, H2O2, and H2O2+ PDGF groups. The cells were pre-treated with PDGF (5 ng/mL) for 2 hours before stimulation with 200 μM of H2O2. The H2O2 concentration was based on our previous research report (Wang et al., 2018). Cell viability was assessed by Cell Counting Kit-8 solution (Beyotime) according to the manufacturer’s protocol. Pericytes (Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd., Shanghai, China) were added to 96-well plates, and the serum was depleted for 24 hours. The cells were then incubated with the indicated concentration of PDGF for 48 hours. Next, 10 μL of Cell Counting Kit-8 solution was added to each well, followed by incubation for 1 hour. The absorbance was measured at a wavelength of 450 nm using a microplate reader (Molecular Devices). Each experiment was performed in triplicate.
5
Cell Viability Assay with CCK-8
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Cell Counting Kit‐8 solution (Beyotime Company, China) was added into 96‐well plates, reaching to 10 μL/well. The 96‐well plates were then cultured at 37°C containing 5% CO2 for 1 hour. The plates were read at 450 nm using a microplate reader (Bio‐Rad).
6
Cytotoxicity Assay for DOX and SML
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The cells were seeded onto 96-well plates at a density of 2,000 cells/well. Following treatment with 1, 0.1 µg/ml DOX for 1, 2, 3 and 4 days; 2, 25, 50, 75 and 100 µg/ml SML for 1, 2, 3 and 4 days; 3, DOX alone or SML plus DOX for 1, 2, 3 and 4 days, the cells were washed with PBS. RPMI-1640 medium (100 µl) and 10 µl cell counting kit-8 solution (Beyotime Institute of Biotechnology) were added to each well for another 2 h at 37°C. The optical density value was read at 450 nm using an ultraviolet spectrophotometer.
7
MCC950 and Quercetin Effects on NLRP3 Inflammasome
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RIMVECs were seeded in a 96-well cell culture plate at a concentration of 1 × 105 cells/well in 100 μL DMEM per well. When the cells reached 80% confluency, they were treated with MCC950 at different concentrations (0, 0.05, 0.5, 5, 10, and 50 μM), according to a previous report (30 (link)) and incubated for 12 h. Cell viability was measured using Cell Counting Kit-8 solution (Beyotime Biotechnology), as explained in the paragraph of cell viability. As regards the viability under different treatments, cells were seeded in a 6-well plate at a concentration of 1 × 105 cells/well in 200 μL DMEM per well. When the cells reached 80% confluency, they were divided into the following groups: control group (no treatment), model group (10 μg/mL LPS 24 h followed by 1 mM ATP 4 h), treatment groups (10 μg/mL LPS simultaneous to quercetin 10 μM 24 h followed by 1 mM ATP 4 h), and MCC950 group (5 μM MCC950 1 h according to the manufacturer’s instructions and then 10 μg/mL LPS 24 h followed by 1 mM ATP 4 h). The protein expression of NLRP3, caspase-1, ASC, GSDMD, ZO-1, claudin-1, claudin-2, IL-1β, IL-18, and IL-6 was evaluated by western blot as explained in the related paragraph.
8
Cell Cycle Assay Protocol
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The cell cycle assay was performed as previously described with a slight
change[18 (link)]. In brief,
cells were seeded into 96-well plates and then harvested (1 x 106cells per group) 48 hours later. After centrifugation for three minutes, the
cells were collected and fixed with 70% ethanol at 4°C. After 0, 24, 48, and 72
hours, 10 µl cell counting kit-8 solution (Beyotime, Shanghai, China) was
added into each well. The cells were cultured for another 0.5 hour before use.
The optical density value was captured by microplate reader (Bio-Rad, Hercules,
California, United States of America) at 450 nm.
change[18 (link)]. In brief,
cells were seeded into 96-well plates and then harvested (1 x 106cells per group) 48 hours later. After centrifugation for three minutes, the
cells were collected and fixed with 70% ethanol at 4°C. After 0, 24, 48, and 72
hours, 10 µl cell counting kit-8 solution (Beyotime, Shanghai, China) was
added into each well. The cells were cultured for another 0.5 hour before use.
The optical density value was captured by microplate reader (Bio-Rad, Hercules,
California, United States of America) at 450 nm.
9
Cell Viability Assay for Transfected SACC Cells
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After transfection as described previously, the SACC-83 and SACC-LM cell lines were seeded into 96-well plates with 5,000 cells/well. A 10-μl quantity of Cell Counting Kit-8 solution (CCK-8; Beyotime Institute of Biotechnology, P.R. China) was added to each well at 0, 24, 48, and 72 h after transfection. The cells were subsequently incubated for 2 h at 37°C. Optical density (OD) was measured at a wavelength of 450 nm.
10
Cell Proliferation Assay for Rat ADSCs
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Rat ADSCs of three passages were seeded into 96-well plates at a cell density of 2×104/ml, and cultured in DMEM supplemented with 10% FBS. After 1, 2, 3, 4, 5, 6 or 7 days' culture, cells were incubated with 10 µl Cell Counting kit-8 solution (Beyotime Institute of Biotechnology, Haimen, China) at 37°C for 3 h. The samples were measured spectrophotometrically at a wavelength of 450 nm with a Microelisa reader (Infinite M200PRO, Tecan Group Ltd., Männedorf, Switzerland).
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