Cell viability was assessed using the cell counting kit-8 (CCK-8, cat#B34302, Bimake, USA) according to the manufacturer’s instructions. Briefly, cells were seeded at a density of 3 × 103 cells per well in 96-well plates in 100 μL culture medium without or with various concentrations of vinblastine and different types of inhibitors, and then incubated in a 37 °C, 5% CO2 incubator. After culturing for 0, 24 or 48 h, 10 μL of the CCK-8 reagent was added to each well and cells were incubated for 2 h at 37 °C. The absorbance of the sample was measured by EL × 800 microplate reader (Biotek Instruments Inc., Vermont, USA.) at 450 nm as previously described [29 (link)]. At least two independent experiments were performed in triplicate.
Cck 8
Manufactured by Selleck Chemicals
Sourced in United States, China
CCK-8 is a colorimetric assay kit used for measuring cell viability and cytotoxicity. It contains a water-soluble tetrazolium salt that is reduced by living cells to produce a colored formazan dye, which can be quantified spectrophotometrically.
Automatically generated - may contain errors
Lab products found in correlation
Elx800, by Agilent Technologies (3 mentions)
Synergy htx, by Agilent Technologies (3 mentions)
Microplate reader, by Bio-Rad (3 mentions)
Prism 7, by GraphPad (2 mentions)
Elx800 microplate reader, by Agilent Technologies (2 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions)
El800 microplate reader, by Agilent Technologies (1 mentions)
Cck 8, by Molecular Devices (1 mentions)
Microplate reader, by Molecular Devices (1 mentions)
Edu incorporation assay, by RiboBio (1 mentions)
Transwell assay, by Corning (1 mentions)
Yefluor 594 edu imaging kit, by Yeasen (1 mentions)
Prism 8, by GraphPad (1 mentions)
Automatic microplate reader, by Bio-Rad (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Gefitinib, by Selleck Chemicals (1 mentions)
Ly294002, by Selleck Chemicals (1 mentions)
Palbociclib, by Selleck Chemicals (1 mentions)
Metformin, by Merck Group (1 mentions)
Cell counting kit 8 (cck8), by Selleck Chemicals (1 mentions)
81 protocols using cck 8
1
Cell Viability Assay Using CCK-8
2
Cell Viability Assay with CCK-8
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The treated cells were seeded in five 96-well plates (1000/well), to one of which CCK-8 was added (Bimake, Houston, TX, USA); incubated in a 37 °C 5% CO2 incubator for 1.5 h; and then measured at 450 nm by using an Eon plate reader, Then, we compared the cell quantity based on the magnitude of the absorbance values to detect cell viability. The rest were also cultured in an incubator, and CCK-8 was added every 24 h to detect one piece.
3
Cell Viability Assay using CCK-8
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The tetrazolium salt CCK-8 (Bimake, China) was used to determine cell viability according to the manufacturer’s protocol. BRL-3A cells were cultured in 96-well plates. After pre-treatment, CCK-8 was added to the cell culture medium at 1:10 and sequentially incubated at 37 °C for 1 h. Finally, absorbance at 450 nm was measured using a microplate reader (Molecular Devices, CA, USA).
4
Cell Viability Assay with MTF1 siRNA
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According to the protocols of cell counting kit-8 (CCK-8, Bimake, B34302), 2 × 104 cells transfected with MTF1 siRNAs were plated into the 96-well plates, and cultured for the indicated times (1 day, 2 day, 3 day, 4 day and 5 day). Each groups contained five replicates. Then, 10 μL CCK-8 reagent was mixed into each well and incubated at 37 °C for about 2 h. The absorption peak intensity of reaction mixture was assessed using an absorbent photometer at 495 nm.
5
Assessing Astrocytoma Cell Viability, Proliferation, and Invasion
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The CCK8 (Bimake) and EDU incorporation assays (Ribobio) were performed according to the manufacturers’ procedures. Cell viability was assessed by a CCK8 assay as previously described (25 (link)). The proliferation ability of astrocytoma cells was detected by an EDU incorporation assay as previously described (26 (link)). The invasive ability of astrocytoma cells was tested by a transwell assay (Corning Inc.) as previously described (25 (link)).
6
Evaluating miR-590-3p Regulation of Cell Proliferation
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1×103cells of different group after transfection (miR-590-3p mimics, NC, inhibitors and inhibitor NC) were seperately seeded into six-well culture plates per well. After 10 days culturing under the condition of 37°C and 5% CO2 in an incubator, the plates were washed with PBS. We washed the cells with PBS twice after fixing them with precooled methanol for 30 min and staining them with 1% crystal violet for 30 min. We used Cell Counting Kit-8 (CCK-8) assay to detect cell proliferation. Briefly, we harvested cells with 0.05% trypsin/EDTA and counted them after transfection (miR-590-3p mimics, NC, inhibitors and inhibitor NC) for 48 h and then seeded them into four 96-well plates at a density of 3000 cells per well and incubated overnight. The cells were added with 10 μL CCK-8 (Bimake; USA) and incubated for three hours at 37°C. The absorbancy value (with OD450 nm) was detected every 24 h for four days by ELISA 96-well microtiter plate reader (BIORAD680; USA) and data are presented as the cell number. Three independent experiments were performed.
7
Cell Proliferation and Viability Assays
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For cell proliferation assay, cells were seeded on a 96-well plate at a density of 1 × 103 cells per well, and CCK-8 assay was performed at 5 time points (0, 24, 48, 72, 96 h) after seeding to evaluate cell proliferation. Cells were incubated in 5% CCK-8 (B34304, Bimake) diluted by DMEM without FBS for 1.5 h before each time point, and the absorbance at 450 nm wavelength was tested to evaluate cell proliferation of each well.
For 5-Ethynyl-2'-deoxyuridine (EdU) staining, cells were seeded on rounded glass cell slides in a 24-well plate for 24 h, then cells were stained using Yefluor 594 Edu imaging Kits (40276ES60, YEASEN) following the manufacturer’s instruction.
For cell viability assay, cells were seeded on a 96-well plate at a density of 1 × 104 cells per well for 24 h, then were treated with or without graded concentrations of erastin for 24 h. Subsequently, cells were incubated in 5% CCK-8 diluted by DMEM without FBS for 1.5 h, and the absorbance at 450 nm wavelength was tested to evaluate cell viability of each well.
For 5-Ethynyl-2'-deoxyuridine (EdU) staining, cells were seeded on rounded glass cell slides in a 24-well plate for 24 h, then cells were stained using Yefluor 594 Edu imaging Kits (40276ES60, YEASEN) following the manufacturer’s instruction.
For cell viability assay, cells were seeded on a 96-well plate at a density of 1 × 104 cells per well for 24 h, then were treated with or without graded concentrations of erastin for 24 h. Subsequently, cells were incubated in 5% CCK-8 diluted by DMEM without FBS for 1.5 h, and the absorbance at 450 nm wavelength was tested to evaluate cell viability of each well.
8
Doxorubicin Cytotoxicity Assessment
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Cell counting kit-8 assay (CCK-8) (Bimake, Houston, TX, USA) was used to monitor the cell viability with its provided protocol. Transfected cells were seeded into 96-well plates at 8 × 103 cells per well. Doxorubicin was added to cells at concentrations of 0, 0.5, 1, 2, 4 and 8 μg/mL, and incubated for 48 h. Then 10 μL/well of CCK-8 reagent was added to the 96-well plates. Two hours later, the absorbance was measured by the microplate reader (ELX800; BioTek, Winooski, VT, USA) at wavelength of 450 nm. The IC50 value was determined by GraphPad Prism 7.0. All experiments were independently repeated three times.
9
Cell Proliferation Assay for NSCLC
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After the cells had reached 90% confluence in culture, the Bease-2b, A549 and NCI-H1299 cells transfected with Si-PLK1 were inoculated into 96-well culture plates at 4000 cells per well, with four replicate wells for each group. They were then subjected to incubation (37°C) in a 5% CO2 incubator and tested for 450 nm absorbance using 10 μL CCK-8 (Bimake, Texas, USA) at 24, 48, and 72 h, and each group of experiments was conducted repeatedly three times.
10
Cytotoxicity Assay of dBET57
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NB cells were divided into 1 × 104 cells, which were inoculated into 96 well plates, incubated overnight in an incubator at 37°C, and then, dBET57 was added to each well according to the concentration gradient. After 72 hours of drug treatment, add CCK8 (B34304, Bimake, TX, USA) reagent to each well based on the instructions. The microplate reader read the absorbance at 450 nm. Each concentration was examined and replicated at least three times independently, then calculated the IC50 values of dBET57 by using GraphPad Prism 8.4.3.
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