Cell Counting Kit-8 (C09-0814-345)

Cell Counting Kit-8(CCK-8)Water-soluble tetrazole (2-(2-methoxy-4nitrobenzene)-3-(4-nitrobenzene)-5-(2,4-disulfobenzene)-2H-tetrazole monosodium salt ), is a compound similar to MTT. In the presence of an electron coupling reagent, it is reduced by dehydrogenase in the mitochondria to an orange-yellow water-soluble formazan dye (formazan, below). This formazan dye Dissolve directly in the medium. The water-soluble tetrazole is bioreduced by intracellular dehydrogenase to produce formazan. The more cells proliferate, the darker the color of the medium; the more cytotoxic, the lighter the color. For the same cell, the number of formazan compounds produced is proportional to the number of living cells, and the color intensity is linearly related to the number of cells. It is the CCK-8 kit developed using this feature to directly analyze cell proliferation and toxicity. The CCK-8 method is widely used and can be used for the activity detection of biologically active factors, the screening of anti-tumor drugs, the determination of cell proliferation, the detection of cytotoxicity, and drug sensitivity and other experiments related to cell viability and proliferation. This kit is easy to use. The kit contains a prepared tube of CCK-8 solution containing water-soluble tetrazole, ready to use, no other preparation steps are required. The detection process does not require additional steps to dissolve formazan. It can be directly detected on a microplate reader using a 96-well plate or a 384-well plate, which is suitable for large-scale high-throughput sample detection.Comparison of CCK-8 method and MTT:The CCK8 kit provides a method for detecting cell proliferation and viability with high sensitivity, simple operation, safe use, and good reproducibility. Compared with traditional MTT, there is no need for organic solvents and radioactive isotopes, fewer steps, no loss, and accurate results! The test kit is very convenient. The kit only has one tube of CCK-8 solution containing WST-8 that has been prepared, and there is no need to perform any preparation operations. There is no need to use isotope, all detection steps are only completed in the same 96-well plate. There is no need to wash the cells, no need to collect the cells, and no additional steps to dissolve formazan. It can be used for the detection of large quantities of samples.1. The formazan produced by the MTT experiment is not water-soluble and needs to be dissolved in organic solvents such as DMSO; while the formazan produced by this method is water-soluble, which not only eliminates the dissolution step, but also reduces the operating steps. Error.2. Compared with the MTT method, this method has a wider linear range and higher sensitivity. This method is non-toxic to cells, so after adding WST-8 for color development, it can be repeatedly measured with a microplate reader at different times to find the best measurement time.3. The reagent used in this method is more stable than MTT in the culture medium, and the experimental effect is reproducible.4. MTT is toxic, and the formazan produced by it needs to be dissolved by organic solvents, which will cause harm to the operators. This reagent is non-toxic, no organic solvent is needed in use, and the operation is safer.5. This kit can be stored for a long time at 4℃ and protected from light. It does not need to be prepared for use, and it is ready to use.6. Phenol red and serum will not interfere with CCK detection (just deduct blank holes)Comparison of the advantages of CCK-8 method and other cell proliferation/toxicity detection methods Detection method  MTT  XTT  WST-1  CCK-8   Water solubility of formazan products  Poor, need to add DMSO to dissolve  Good  Good  Good   Product traits  Powder  2 bottles of solution  Solution  1 bottle of solution   Instructions  Use after mixing  Should be prepared when using   Ready to use  Ready to use   Detection sensitivity  Generally  Higher  Higher  high   Reproducibility  Difference  Medium  Good  Very good   Detection time  Longer  Shorter  Shorter  Shortest   Detection wavelength  560-600nm  420-480nm  420-480nm  430-490nm   Cytotoxicity  High  Very low  Very low  Very low   Cell morphology disappeared completely  Unchanged cell morphology  Unchanged cell morphology  Unchanged cell morphology   Reagent stability  Generally  Very bad  Generally  Well   Batch sample inspection  Can  Very suitable  Very suitable  Very suitable   Convenience  Generally, heavy workload  Convenient  Convenient  Very convenient Packing: 1ml/100TStorage conditions: CCK-8 solution can be stored for one year under the conditions of dark and 0-5℃; 2 years at -20℃; if you need to use it frequently, please store the reagent at 0-5℃ to prevent the background value from increasing Do not freeze and thaw repeatedly to interfere with the experimental results.light & Moisture sensitive. 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