The diluted suspensions (2 ml), containing different cell densities, were added into the wells of 24-well plates and cultured for 24 h. Specifically, the number of cells in each well was 0, 500, 1,000, 2,000, 5,000, 10,000, 20,000, and 40,000. Discarding the medium and cleaning three times with PBS, 450 µl medium and 50 µl CCK8 kit (BestBio, Shanghai, China) solution were added into the wells and then incubated at 37°C for another 2 h. Then, the culture medium was mixed thoroughly, and the supernatant with a volume of 100 µl was taken to a 96-well plate to measure the absorbance at 450 nm with a microplate reader (BL340, Biotech, United States). With the number of cells as the abscissa and the absorbance value as the ordinate, the standard curve was drawn and the relationship between the number of cells and the absorbance value was obtained by linear regression fitting. CCK8 test of the cells on the scaffolds was repeated every 24 h for 5 days. The number of cells on the scaffold with five different SLS parameters at each selected time point (days 1, 2, 3, 4, and 5 after cell seeding) was calculated according to the obtained equation.
Cck 8 kit
Manufactured by BestBio
Sourced in China
The CCK-8 kit is a colorimetric assay used to measure cell viability and cytotoxicity. It utilizes the water-soluble tetrazolium salt WST-8 to produce a water-soluble formazan dye upon reduction by dehydrogenases in viable cells. The absorbance of the formazan dye is directly proportional to the number of living cells.
Automatically generated - may contain errors
Lab products found in correlation
Everolimus, by Apexbio (1 mentions)
Elx808 absorbance reader, by Agilent Technologies (1 mentions)
Imark microplate reader, by Bio-Rad (1 mentions)
Ultraviolet spectrophotometer, by Eppendorf (1 mentions)
Varioskan flash spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Harvard Bioscience (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
Victor nivo, by PerkinElmer (1 mentions)
17 protocols using cck 8 kit
1
Cell Viability Evaluation on Scaffolds
2
Podocyte Viability Assay with QTXZG Serum
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Cell viability was determined via a CCK-8 kit (Bestbio, Shanghai, China) and titrated for the optimal time and serum concentration. A 100 µL glomerular podocyte suspension was inoculated into 96 well plates at a cell density of 5 × 104 cells/well for 12 h at 37 °C in a 5% CO2-saturated humidity incubator. Cells were divided into two groups: control group (cell, PAN) and control + QTXZG serum (cell, PAN, and 1.25%, 2.5%, 5%, 10%, 20%, or 40% serum QTXZG). Cells were removed 0, 12, 24, 48, or 72 h after treatment, and 10 μL of CCK-8 reagent was added to each well, after which they were placed in an incubator for 1 h and removed, and the OD value of each well was measured using a microplate analyzer at a wavelength of 450 nm. The cell viability rate was calculated as follows:
Cell viability rate (%) = [ODdrug - ODblank]/[ODcontrol - ODblank] × 100%
Cell viability rate (%) = [ODdrug - ODblank]/[ODcontrol - ODblank] × 100%
3
Cell Proliferation Assay Using CCK-8
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The Cell Counting Kit-8 (CCK-8) was employed to analyse cell proliferation, as described previously25 (link). MCs were plated at a density of 5,000 cells/well in 96-well plates, and subsequently transfected with the miR-27a inhibitor, PPARγ siRNA, or negative controls at a final concentration of 50 nM. At 48 h after transfection, cell proliferation was measured with the CCK-8 Kit (BestBio, Shanghai, PRC). Each assay was performed with 6 replicates in 3 independent experiments.
4
Evaluating Everolimus Efficacy in METTL3 Models
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For the in vitro study, everolimus (APExBIO, Houston, TX, USA) was added to cells with final concentrations of 5 or 50 µg/mL. Cell viability was measured using a CCK-8 kit (BestBio, Shanghai, China). For the in vivo study, subcutaneous xenograft models with METTL3-high (n = 3) and control cells (n = 3) were established as above. Control cells were inoculated two days before METTL3-high cells. When the tumor sizes were similar, volume-matched mice received everolimus (50 µg/day intragastrically) or solvent for 17 days and were sacrificed at day 18. Mice were randomly allocated to each group with no blinding.
5
Cytotoxicity Evaluation of Tan IIA
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Cell viability was assessed with the CCK-8 Kit (Bestbio) following the manufacturer's instructions. Briefly, 1×106 cells were seeded in 100 µl culture medium on 96-well plates and then treated with different concentrations of Tan IIA (0.25, 0.5, 1, 2, 4, 8, 16 and 32 µM). After 48 h, 10 µl CCK-8 assay solution was added to each well and the cells were further incubated at 37°C with 5% CO2 for 2 h. The optical density values were measured using an ELx808 absorbance reader (BioTek Instruments, Inc.) at 450 nm.
6
Cell Viability Measurement by CCK-8
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The cell viability was measured by the CCK-8 kit (Bestbio, China Co., Ltd.) and determined for 4 days. The cells after 72 hours of infection were seeded in a 96-well plate at about 6.0x103 cells/well. 10μl of CCK-8 solution was added to the medium, and the Y14 cell was incubated at 37°C for 2 hours. The proliferation rate of Y14 cell was detected at 0, 24, 48, 72, and 96 after infection. Optical density (OD) value was measured at 450 nm (iMarkTM Microplate Reader, Bio-Rad, USA). All of these experiments were repeated three times.
7
PRMT5 Knockdown Impacts Cell Proliferation
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A cell counting kit (CCK)‐8 kit (BestBio, Shanghai, PR China) was used to determine the effect of PRMT5 knockdown on cell proliferation function. The transfected cells were placed into 96‐well plates, with 2000 cells in 100 μl of culture medium per well. Next, 10 μl of the CCK‐8 reagent was added at 0, 24, 48, 72, and 96 h, followed by further incubation at 37 °C for 90 min. The absorbance was measured using an ultraviolet spectrophotometer (Eppendorf, Hamburg, Germany) with the wavelength set at 450 nm. Three auxiliary wells were set for each assay, and the assays were repeated thrice.
8
Fucoidan Cytotoxicity Evaluation
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Based on the instructions of the CCK8 kit (Best Bio, Shanghai, China), we digested cells in logarithmic growth phase into a cell suspension and counted them. Then, 2000 cells were inoculated per well of a 96-well plate. Each 96-well plate contained 100 μl of different concentrations of fucoidan. Later, the proliferation ability of A549 and H1650 cells was analyzed at 24, 48, 72 and 96 h. Then, 100 μl of fresh medium containing 10 μl of CCK8 solution was added to each well and cultivated for 2 h. We measured the absorbance at 450 nm with a Thermo Scientific Varioskan Flash spectrophotometer (Thermo Scientific, Inc., Vantaa, Finland).
9
Cell Viability Assay of U0126 Treatment
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CCK-8 kit (BestBio, China) was used to detect the cell viability (Lv et al., 2018 (link)). In brief, CEF cells in 96-well plates were treated with 20 μM U0126 dissolved in DMSO for 24 and 48 h. Then, 10 μl CCK-8 reagent was added into the cells and incubated in the dark for 2 h at 37°C. A spectrophotometer (Bio-Rad) was used to read the OD450 of each well. Viabilities of the CEF cells treated with U0126 were compared with that of negative control.
10
Doxorubicin-Induced LY8 Cell Proliferation
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LY8 cells treated with 10, 20, and 40 nM doxorubicin, respectively, six times were plated at a density of 5000 cells/well. Cell proliferation was measured with a CCK‐8 Kit (BestBio). Each assay was performed with five replicates in three independent experiments.
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