Enhanced CCK-8 is a reagent kit based on WST-8 that is widely used for rapid and highly sensitive detection for cell proliferation and cytotoxicity. It has better detection sensitivity and a wider linear range than ordinary CCK-8 reagent kits.
WST-8 is a compound similar to MTT, which can be reduced by some dehydrogenases in the mitochondria to form orange formazan in the presence of an electron-coupling reagent. The more rapidly cells proliferate, the deeper the color, and the greater the cytotoxicity, the lighter the color. For the same cells, the depth of color is linearly related to the number of cells.
WST-8 VS MTT
WST-8 is an upgraded alternative to MTT, and has clear advantages over MTT and other similar products such as XTT and MTS. Firstly, the formazan produced by the reduction of MTT by some dehydrogenases in the mitochondria is not water-soluble, requiring a specific solvent for dissolution; whereas the formazan produced by WST-8, XTT, and MTS are all water-soluble, eliminating the need for subsequent dissolution steps. Secondly, the formazan produced by WST-8 is more easily soluble than that produced by XTT and MTS. Thirdly, WST-8 is more stable than XTT and MTS, resulting in more stable experimental results. In addition, the linear range and sensitivity of WST-8 are wider and higher than those of MTT, XTT, and other similar products.
Product Advantage
This reagent kit is very convenient to use. The kit contains a single tube of pre-formulated Enhanced CCK-8 solution containing WST-8, with no need for any additional preparation or operations. Isotopes are not required, and all detection steps are completed in the same 96-well plate. There is no need to wash cells, collect cells, or use additional steps to dissolve formazan. It can be used for testing large quantities of samples.
Note
1.Due to the use of a 96-well plate for detection, if the cell culture time is prolonged, evaporation must be taken into account. On the one hand, since the outermost rim of the 96-well plate is the most prone to evaporation, one solution is to abandon that rim and add the same amount of PBS, water, or culture medium. On the other hand, the 96-well plate can be placed near a water source in the incubator to mitigate evaporation.
2.The detection in this reagent kit depends on the reaction catalyzed by dehydrogenase, so reducing agents (such as some antioxidants) can interfere with the detection. If there are a large amount of reducing agents in the system being tested, they must be removed.
3.Before using an ELISA reader for detection, make sure there are no air bubbles in each well, as this will interfere with the assay.