introduction
Cell Counting Kit-8 (abbreviated as CCK-8) is a detection reagent widely used for cell proliferation and cytotoxicity assays based on WST-8. WST-8 [chemical name: 2-(2-Methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt] is a compound similar to MTT. In the presence of the electron carrier 1-Methoxy PMS (1-Methoxy-5-methylphenazinium methyl sulfate), it is reduced by dehydrogenases in mitochondria to a highly water-soluble orange-yellow formazan product.
The more and faster the cells proliferate, the darker the color will be; the greater the cytotoxicity is, the lighter the color will be. For the same type of cells, the depth of the color is proportional to the number of living cells. Therefore, this characteristic can be utilized to directly conduct analyses on cell proliferation and cytotoxicity.
CCK8 Assay VS MTT Assay
A. The MTT assay has the maximum absorbance value at 570 nm, and its linear range is around 1. Moreover, the MTT dye is lipid-soluble. Therefore, when it comes to the final color development, an organic solvent like DMSO is required to dissolve the dye. It has certain toxicity and complicates the experimental procedure.
B. The CCK8 assay has the maximum absorbance value at 450 nm, and it has a wider linear range. A value not exceeding 3 is acceptable. Additionally, the CCK8 dye is water-soluble, making the experimental operation simple and less toxic.
advantages
✔ Provide the results of cell proliferation at four time points: 0 h, 24 h, 48 h, and 72 h. Ensure that the initial amount of cells input is consistent and that there is a consistent trend at different time points, so as to obtain more reliable phenotypic results.
✔ Data analysis, graphing, and English materials and methods, as well as detailed raw data, will safeguard your article publication.
✔ Examples of published articles.
workflow
Delivery Contents:
A. Raw data of all experimental results.
B. Experimental reports (including detailed experimental procedures and the manufacturer and catalog number information of the main instruments and reagents).
C. English materials and methods.
demo data
