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Storage Store at 4℃ and protect from light. Long-term storage can be placed at -20℃. Expiration date marked on the outer packing.
- Cell Counting Kit-8, based on WST-8 (chemical name: 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfob enzene)- 2H-tetrazole monosodium salt), is a rapid and highly sensitive detection kit widely used in cell proliferation and cytotoxicity. In the presence of electron coupling reagent, WST-8 can be reduced by dehydrogenase in mitochondria to form orange formazan, which is highly water-soluble.
- The depth of color is directly proportional to cell proliferation and inversely proportional to cell toxicity. OD value was measured at 450 nm wavelength by enzyme labeling instrument, which indirectly reflected the number of living cells.
- WST-8 is an upgrade substitute product of MTT, which has obvious advantages compared with MTT or other similar products such as XTT or MTS. First of all, formazan, which is reduced by some dehydrogenase in mitochondria, is not water-soluble and needs to be dissolved in a specific solution; while formazan, produced by WST-8, XTT and MTS, is water-soluble, so subsequent dissolution steps can be omitted. Secondly, formazan produced by WST-8 is more soluble than that produced by XTT and MTS. Thirdly, WST-8 is more stablethan XTT and MTS, which makes the experimental results more stable.
- In addition, WST-8 has wider linear range and higher sensitivity than MTT and XTT.
Method of application
Protocol Determination of cell activity
- Cell suspension (100 μL/well) was inoculated into 96 well plates. Put the culture plate in the incubator for preculture (37℃, 5% CO2).
- Add 10 μL CCK solution to each hole (be careful not to generate bubbles in the well, they will affect the OD value). Note: If the liquid is not clarified or precipitated after taking it out of the refrigerator, it can be used after 37°C water bath treatment without affecting the result.
- Incubation at 37℃,5% CO2 for 1-4 h.
- Determination the absorbance at 450 nm by enzyme scale.
- If OD value is not determined temporarily, 10 μL 0.1 M HCl solution or 1% w / v SDS solution can be added into each well, and the culture plate shall be covered and stored at room temperature in dark. The absorbance will not change within 24 hours. Determination of cell proliferation/ cytotoxicity .
Cell Counting Kit-8(CCK-8)
|Component||1 mL||5 mL||30 mL||100 mL||5×100 mL|
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