CCK-8 Data Analysis Tutorial: A Comprehensive Guide238

The Cell Counting Kit-8 (CCK-8) assay is a commonly used method for determining the viability of cells in response to various treatments or conditions. CCK-8 is a tetrazolium salt that is reduced to a colored formazan product by metabolically active cells. The absorbance of the formazan product can be measured using a microplate reader to quantify the number of viable cells.

Materials Required* CCK-8 assay kit
* Cells
* Culture medium
* Microplates
* Microplate reader

Procedure1. Prepare cell suspensions in culture medium according to the manufacturer's instructions.
2. Add 100 μL of cell suspension to each well of a 96-well microplate.
3. Add 10 μL of CCK-8 solution to each well.
4. Incubate the microplate for 1-4 hours at 37°C in a humidified incubator.
5. Measure the absorbance of the formazan product at 450 nm using a microplate reader.

Data AnalysisThe absorbance values obtained from the CCK-8 assay can be used to calculate the percentage of viable cells in each sample. The following formula can be used:
Percentage of viable cells = (Absorbance of sample / Absorbance of control) x 100
The control is typically a sample of untreated cells.
The data can be plotted as a bar graph or a line graph to visualize the effects of different treatments or conditions on cell viability.

Statistical AnalysisThe statistical significance of differences between groups can be determined using a one-way analysis of variance (ANOVA). If the ANOVA indicates that there is a significant difference between groups, a post-hoc test can be used to determine which groups are significantly different from each other.

Troubleshooting* Low absorbance values: This can be due to insufficient incubation time, low cell density, or low metabolic activity.
* High absorbance values: This can be due to over-incubation, high cell density, or non-specific binding of the formazan product.
* Variable absorbance values: This can be due to pipetting errors or differences in cell growth.

ConclusionThe CCK-8 assay is a simple and convenient method for determining the viability of cells. The data obtained from the assay can be used to assess the effects of different treatments or conditions on cell growth and survival.

2025-02-11

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