1. Kit preparation.
Before manipulation, make sure that the CCK8 kit has been removed from the freezer and placed at room temperature to allow it to return to room temperature. When you open the kit, you can see that it contains components such as CCK8 solution, wash solution, and stop solution.
2. Cell seeding.
1.According to the experimental requirements, the cells are seeded in 96-well plates. In general, 100-200 ul of cell suspension is added to each well and placed in an incubator with %CO2 for culture.
2.When the cells grow to the desired density, different concentrations of drug solutions are added and the culture is continued for a certain period of time.
3.Cell growth was observed at different time points after dosing, and data were recorded.
3. Reagent preparation.
1.Prepare components such as CCK8 solution, wash solution, and stop solution according to the instructions on the instructions.
2.Aliquot the prepared solution into 96-well plates and add 100 ul per well.
3.After adding the CCK8 solution, gently shake the 96-well plate to mix well.
4. Colorimetric determination.
1.Remove the 96-well plate at the set assay time point and add the stop solution to the 100ul wells.
2.The OD value of each well was determined by a microplate reader, and the detection wavelength was 450 nm.
3.Cell viability is calculated based on the measured OD values, and cell growth curves are plotted.
5. Analysis of results.
1.Calculate cell growth inhibition rate or cell viability rate based on experimental data.
2.Experimental data were compared with the control group to analyze the effect of the drug on cell growth.
3.Visualize cell growth inhibition or survival curves to assess the cytotoxic effects of drugs.
6. Precautions.
1.During the operation, be sure to follow the steps on the instructions to avoid affecting the accuracy of the experimental results.
2.Do not use expired reagents and instruments to avoid affecting the experimental results.
3.When using reagents and equipment, please pay attention to safety precautions, such as wearing gloves and avoiding direct contact**.