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Cell Counting Kit-8 (CCK-8)
The CCK-8 assay kit is a highly sensitive kit based on WST-8, widely used for rapid detection of cell proliferation and cell toxicity, and can be used as a perfect alternative to the MTT assay. WST-8 is reduced by dehydrogenases in mitochondria to form orange-colored formazan product in the presence of an electron coupling agent (1-Methoxy PMS), and the amount of formazan generated is directly proportional to the number of viable cells. Therefore, this characteristic can be utilized for cell proliferation assays, toxicity analysis, drug screening, tumor drug sensitivity tests, and other applications.
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Cat. No.: CCK-1 (for 1ml)
Cat. No.: CCK-5 (for 5ml)
Cat. No.: CCK-25 (for 25ml)
Description
The CCK-8 assay kit is a highly sensitive kit based on WST-8, widely used for rapid detection of cell proliferation and cell toxicity, and can be used as a perfect alternative to the MTT assay. WST-8 is reduced by dehydrogenases in mitochondria to form orange-colored formazan product in the presence of an electron coupling agent (1-Methoxy PMS), and the amount of formazan generated is directly proportional to the number of viable cells. Therefore, this characteristic can be utilized for cell proliferation assays, toxicity analysis, drug screening, tumor drug sensitivity tests, and other applications.
The CCK-8 assay kit has the following advantages:
- High sensitivity and good reproducibility.
- Easy to use, ready-to-use, and convenient.
- Reagents are water-soluble, no need for medium exchange, making it particularly suitable for suspension cells.
- Low toxicity to cells, with minimal impact on cell viability.
- Suitable for high-throughput drug screening and other large-scale experimental applications.
Protocol
- Prepare a cell suspension and perform cell counting.
- Seed the cell suspension into a 96-well plate, 100 μl per well, with approximately 2000-5000 cells per well (the specific number of cells used per well should be determined based on the experiment), with 2-3 replicate wells for each sample.
- Incubate the plate in a 37°C cell culture incubator for 2-4 hours to allow the cells to adhere, or overnight for suspension cells.
- Add 10 μl of CCK-8 solution to each well, and gently tap the plate to ensure thorough mixing.
- Incubate the plate in a 37°C cell culture incubator for 1-4 hours. The optimal incubation time for Formazan formation may vary depending on the specific cell type.
- Measure the absorbance at 450 nm using a microplate reader. It is recommended to use a dual-wavelength mode, with a detection wavelength of 450-490 nm and a reference wavelength of 600-650 nm.
Additional Notes
- If the absorbance measurement cannot be performed immediately, 10 μl of 0.1 M HCl solution or 1% w/v SDS solution can be added to each well, and the plate should be covered and stored at room temperature in the dark. The absorbance should be measured within 24 hours, as absorbance may change over time.
- If the drug to be tested has oxidizing or reducing properties, it is recommended to replace the medium with fresh medium before adding the CCK-8 solution to exclude the influence of the drug. If the drug has minimal effects, an equal amount of the drug can be added to the medium as a blank control to minimize the impact.
- During incubation in the cell culture incubator, the outermost wells of the plate are more prone to drying and evaporation, which can result in inaccurate volumes and increased errors. To avoid this, only add medium or PBS to the outermost wells of the plate.
- Bubbles may interfere with the measurement results, so it is important to ensure that there are no bubbles in the wells before measuring with a microplate reader.
Storage
The CCK-8 assay kit can be stored at 4°C, with a shelf life of 1 year. Alternatively, it can be stored at -20°C, with a shelf life of 2 years. Please store the kit in a light-protected manner.
Related:
- Cell Counting Kit-8 (CCK-8)
- Enhanced Cell Counting Kit-8
- Super-Enhanced Cell Counting Kit-8
- Cell Counting Kit-3D
Only for research and not intended for treatment of humans or animals
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