Cell viability of PDLCs was estimated by Cell Counting Kit-8 (CCK8) assay and the whole procedure was carried out following the instructions of Cell Counting Kit-8 (Beyotime). 96-well plates of PDLCs were incubated with 10 μL of CCK8 solution 24 h after transfection and then the absorbance at 450 nm was measured.
Cell counting kit 8 (cck8)
Manufactured by Beyotime
Sourced in China, United States, Puerto Rico
The Cell Counting Kit-8 is a colorimetric assay used to measure the number of viable cells in a sample. It utilizes a water-soluble tetrazolium salt that is reduced by cellular dehydrogenase enzymes to produce a colored formazan dye, which can be measured using a spectrophotometer.
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Lab products found in correlation
Microplate reader, by Bio-Rad (88 mentions)
Microplate reader, by Thermo Fisher Scientific (88 mentions)
Microplate reader, by Agilent Technologies (46 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (26 mentions)
Multiskan fc, by Thermo Fisher Scientific (19 mentions)
Elx800, by Agilent Technologies (18 mentions)
Spectramax m5, by Molecular Devices (17 mentions)
96 well plate, by Corning (16 mentions)
Microplate reader, by Molecular Devices (10 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (10 mentions)
Multiskan go, by Thermo Fisher Scientific (10 mentions)
Multiskan mk3, by Thermo Fisher Scientific (8 mentions)
Cck 8, by Beyotime (8 mentions)
Varioskan flash, by Thermo Fisher Scientific (8 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (8 mentions)
Microplate reader, by Tecan (7 mentions)
96 well microplate, by Corning (7 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (7 mentions)
Trypsin edta, by Thermo Fisher Scientific (7 mentions)
Infinite m200, by Tecan (6 mentions)
1 259 protocols using cell counting kit 8 (cck8)
1
Cell Viability Assay of PDLCs
2
Evaluating Cell Viability with DA Treatment
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RaECs or HMEC‐1 cell lines were seeded into 96‐well plates at a density of 1 × 104 cells per well (Corning) under the above conditions for 24 h prior to the experiment. The culture medium was replaced with 100 μL of DA solutions at different concentrations. Each concentration was replicated in five wells. The cells were incubated for another 24 h. PBS was used to clean each hole and then replaced it with freshly prepared serum‐free medium with 10% cell counting kit‐8 (Beyotime), and the cells were incubated for another 1–4 h. The optical density readings were performed using a multimode plate reader (Bioreader) at a wavelength of 450 nm. The absorbance was read relative to the blank well. Cell viability (%) in each well was calculated by (OD450 test – OD450 blank)/(OD450 control – OD450 blank) × 100%. A non‐toxic dose was used to evaluate the effects of DA on RaECs or HMEC‐1 cells.
3
Microglia Viability Assay with RUX
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Microglia (1 × 104/well) were seeded in a 96-well plate and then with a concentration gradient of RUX (0.1–5 mM) for 24 hours. Then, the cells were washed twice and incubated with 100 μL Dulbecco's modified Eagle medium containing 10% cell counting kit-8 (Cat# C0038; Beyotime, Shanghai, China) buffer for 2 hours at 37°C. Detection was carried out at 450 nm using a microplate reader (BioTek, Friedrichshall, Germany), and analysis was performed using ImageJ software (Ver. 1.3.8) (Schneider et al., 2012).
4
Cell Viability Assay with CCK-8
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Cells were seeded in 96-well plates (5 × 103 cells/well). After drug treatment at the indicated doses for 48 h, cell viability were measured using the Cell Counting Kit-8 (CCK8, Biyuntian, Beijing, China) assay. Briefly, discard the medium, add diluted CCK-8 solution (10% in medium) to each well and incubated further for 1.5 h. The optical density value was measured at a wavelength of 450 nm. The following formula was used to calculate the cell survival rate: Cell survival rate (%) = [(As-Ab) / (Ac-Ab)] × 100%.
5
Cell Proliferation Assay Protocol
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A total of 3 × 103 cells were cultivated in 96-well plates, and the culture solution was replaced with fresh RPMI 1640 every day. The Cell Counting Kit-8 (BiYunTian, China) was used to test cell proliferation after 24, 48, and 72 h of incubation.
6
Cell Proliferation Assay Protocol
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After the 24 h treatment period, cells suspensions of exposed and sham-exposed groups were washed by centrifugation to remove trypsin, and then cells were resuspended in fresh DMEM medium. Cells suspension was then seeded into five pieces of 96-well dishes. In each dish, eight wells were seeded for each of the two groups (1000 cells/100 µL/well). Following culture for 24 h, 10 µL of the CCK-8 solution (Cell CountingKit-8, Bi Yun Tian, Nanjing, China) were added to each well and further incubated for 2 h, according to the manufacturer’s directions. The CCK-8 kit is based on the MTT assay, which relies on the transformation of MTT to form a non-water-soluble dye by the dehydrogenase activity within viable cells. Hence, the colorimetric quantification, which is performed at 450 nm using a spectrophotometer (Bio-Rad 680, Bio-Rad, Hercules, CA, USA), provides optical density values that correlate with the proportion of proliferating cells. Values averaging the optical density readings of five dishes for the exposure and sham-exposure group were used to make a proliferation curve.
7
Viability of Transfected Liver Cells
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Transfected cancerous liver cells were seeded into 96-well plates at a density of 1×104 cells per well. Cell Counting Kit-8 (Biyuntian biotechnology) reagent of 10 uL was added to each well after 0 and 2 h. Cell viability of the intervened cells was detected using a microplate reader (DiaTek) at 450 nm absorbance.
8
Sodium Tellurite Oxidative Stress Assay
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Sodium tellurite (Na2TeO3, 99.9%), reduced GSH (98%) and Tf were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Hydroxychloroquine sulfate (HCQ·H2SO4, 99%) was acquired from Adamas-Beta Reagents Co., Ltd. (Shanghai, China). Sodium hydroxide (NaOH, 95%), Sodium chloride (NaCl, 99.5%), and Ethylacetate (99%) were gained from Shanghai Yi En Chemical Technology Co., Ltd. (Shanghai, China). The sulfhydryl assay kit was bought from Solarbio Science & Technology Co., Ltd. (Beijing, China). Fluorescein isothiocyanate (FITC, ≥95%) was obtained from Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Rabbit anti-LC3B and rabbit anti-SQSTM1/p62 antibodies were obtained from Beijing Boosen Biotechnology Co., Ltd. (Beijing, China). The reactive oxygen species detection kit and Cell Counting Kit-8 were obtained from Biyuntian Bio-Technology Co., Ltd. (Shanghai, China). Rabbit IgG/Cy3 was obtained from Wuhan Xavier Biotechnology Co., Ltd. (Wuhan, China). Calcein-AM/PI double stain kit was bought from Yi Sheng Biotechnology Shanghai Co., Ltd. (Shanghai, China).
9
Cell Viability Assay of Mouse Fibroblasts
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Cell viability was assessed using the Cell Counting Kit-8 (CCK-8, Biyuntian Biotechnology, Jiangsu, China) assay. Mouse skin fibroblasts were seeded into 96-well microplates at a density of 5 × 103 cells/well after electroporation. Cell viability was assessed by incubating each well with 100 µL of CCK-8 solution for 4 h after 48 h of culture under the designated conditions (37 °C and 5% CO2), and the absorbance at 450 nm was measured.
10
Cell Viability Assay with CCK-8
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After 48 h co-treatment of AA and LPS, The number of cells was determined using a cell counting kit-8 (biyuntian, Nantong, China). 10 μl of CCK-8 was added into medium directly and then the plates were incubated for an 2 h, and the absorbance at 450 nm was measured using a microplate reader (Molecular Devices, Sunnyvale, CA, USA).
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