The same culture conditions as previously were used to determine whether cells were viable using a cell-counting kit (CCK8 kit, abs50003-5ml, Absin, Shanghai, China). The experiment consisted of four groups: a control group (cells + medium), a blank group (medium), and a group with different drug delivery concentrations (cells + medium + 10/20 μM Cant). Cells were counted, adjusted to a concentration of 2 × 105 mL, and planted at 100 μL/well in 96-well plates using cells from the various treatment groups. Three copies of the cells from each treatment group were planted. The plates were placed in an incubator (37 °C and 5% CO2) and the cells were incubated until the appropriate time. Then, the drug for the pair was added to the corresponding wells at the set concentration. CCK8 solution (10 μL) was added to each well. Then, the cell culture plates were allowed to sit in the incubator for one to four hours. Finally, a plate reader was used to measure the absorbance at 450 nm. Untreated cells, medium, and CCK8 solution were also tested as control wells.
Cck8 kit
Manufactured by Absin
Sourced in China
The CCK8 kit is a cell counting and cytotoxicity assay used to determine the viability and proliferation of cells in culture. It measures the metabolic activity of cells, which is proportional to the number of viable cells. The kit utilizes a water-soluble tetrazolium salt that is reduced by the cellular dehydrogenase enzymes to form a colored formazan product, which can be detected and quantified using a spectrophotometer or a microplate reader.
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Lab products found in correlation
5 protocols using cck8 kit
1
Cell Viability Assay with Cantharidic Acid
2
Quantifying HL-1 Cell Proliferation
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The CCK-8 kit (#abs50003-5 ml, absin) was used per the manufacturer's instructions to detect the proliferation of HL-1 cells following various treatments. Briefly, HL-1 cells were digested using Trypsin-EDTA (0.25%, #abs47047375, absin) and then plated at a density of 5 × 103 cells per well in a 96-well plate. After 24h, 48h, and 72 h, CCK-8 was added to detect the proliferation of HL-1 cells via Elx800 at 450 nm spectrophotometry.
3
Cell Proliferation and Invasion Assays
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The CCK8 kit (Absin; abs50003) was used to measure the cell proliferation levels. The cell invasion assays were conducted in triplicate using transwell chambers (8 μm pore size; Millipore) and chambers coated with Matrigel,19 (link) respectively. The details can be seen in Supplementary Methods (http://links.lww.com/HC9/A14 ).
4
Cell Viability Evaluation by CCK-8 Assay
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We used the CCK-8 kit (abs50003, absin, China) to assess cell viability. Based on the above groups, cells (1 × 104 per well) were transfected and then maintained in an incubator for a specified time (24 or 48 h). Next, 10 μl CCK-8 solution was adopted to treat cells for 4 h. Then, the optical density at a wavelength of 450 nm was determined under a microplate reader (Z742711, Sigma, USA).
5
Evaluating AS-IV Cytotoxicity in PAM Cells
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PAM cells were cultured in 96-well plates at a density of 5 × 103 cells/well, and, after incubation for 24 h, the cells were treated with different concentrations (200, 100, 50, 25, 12.5, and 6.25 μg/mL) of AS-IV (Chengdu Must Bio-Technology Co. Ltd., Chengdu, China) cultured at 5% CO2, 37 °C for 48 h. Cell viability was assessed by employing a CCK8 kit (Absin, Shanghai, China) per the manufacturer’s guidelines.
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