Description

Description: Introduction Elabscience® Enhanced Cell Counting Kit 8 (WST-8 / CCK8) is a rapid, highly sensitive, non-radioactive colorimetric test kit based on WST-8 and widely used in the detection of cell proliferation and cytotoxicity. WST-8 is an analog of MTT, an upgraded replacement product of MTT. Compared with MTT, WST-8 has obvious advantages. First of all, the formazan produced after the reaction is water-soluble and does not require a specific solvent to dissolve it. Secondly, WST-8 is more stable, has a wider linear range and higher sensitivity. Detection principle WST-8 is a compound similar to MTT, which can be reduced to orange formazan by some dehydrogenase in mitochondria in the presence of electron coupling reagent. The amount of formazan produced is directly proportional to the number of living cells. By measuring the absorbance at 450 nm, the amount of living cells can be calculated indirectly. Components Storage Store at 2~8°C for one year in dark or at -20°C for two years. Cautions 1. This kit is for research use only. 2. For your safety and health, please take safety precautions and follow the procedures of laboratory reagent operation. Wear laboratory clothes and disposable gloves during operation, and avoid direct contact with the human body or inhalation of the body. 3. For long time storage, please store at -20°C. For ordinary usage, please store at 2~8°C. Avoid freeze / thaw cycles. 4. Pay attention to mixing during cell seeding to avoid unequal number of cells per well due to cell sedimentation. 5. The incubation time of CCK-8 is generally 1-4 hours. It is recommended to take a preliminary experiment to explore the optimal number of cells and the incubation time of CCK-8. 6. CCK-8 has very low toxicity to cells. Because the dehydrogenase in living cells is continuously produced, CCK-8 can continuously react with the dehydrogenase in living cells. So the color of the solution will be darker and the OD value will continue to increase. 7. The phenol red in the medium will not affect the experimental results. The absorbance of phenol red can be eliminated by subtracting the background absorbance in the blank well during calculation, so it will not affect the detection. 8. When using a 96-well plate for cell culture, pay attention to the result error caused by water evaporation. It is recommended to discard the outer circle of wells and add PBS, water or cell culture medium to prevent water evaporation. In addition, the 96-well plate can also be placed in the incubator near the water source. 9. In order to improve the accuracy of results, make sure that there is no bubble in each well when measuring the OD value with the microplate reader, otherwise it will interfere with the determination. In addition, it is recommended to use a multi-channel pipette to reduce the difference between parallel wells. 10. The detection of this kit relies on the dehydrogenase catalyzed reaction, so reducing agents (such as some antioxidants) will interfere with the detection. If there are many reducing agents in the system to be detected, try to remove them. Or replace the fresh medium before adding CCK-8 to remove the influence of the drug to be tested. 11. If the added medicine contains metal, it will affect the color development. The final concentration of 1 mM ferrous chloride, ferric chloride, and copper sulfate will inhibit 5%, 15%, and 90% of the color reaction and reduce the sensitivity. If the final concentration is 10 mM, it will be 100% inhibited.

Additional Text

-20°C, shading light