Dojindo Cell Counting Kit-8 CCK-8細(xì)胞計(jì)數(shù)試劑盒檢測方法簡介

CCK-8 Kit 細(xì)胞活性檢測試劑中含有WST-8，化學(xué)名：2-（2-甲氧基-4-硝基苯基）-3-（4-硝基苯基） -5-（2,4-二磺酸苯）-2H-四唑單鈉鹽]，它在電子載體1-甲氧基-5-甲基吩嗪鎓*（1-Methoxy PMS）的作用下被細(xì)胞線粒體中的脫氫酶還原為具有高度 水溶性的黃色甲瓚產(chǎn)物（Formazan）。生成的甲瓚物的數(shù)量與活細(xì)胞的數(shù)量成正比。用酶聯(lián)免疫檢測儀在450nm波長處測定其光吸收值，可間接反映活 細(xì)胞數(shù)量。該CCK-8 Kit方法已被廣泛用于一些生物活性因子的活性檢測、大規(guī)模的抗腫瘤藥物篩選、細(xì)胞增殖試驗(yàn)、細(xì)胞毒性試驗(yàn)以及藥敏試驗(yàn)等。
 

Dojindo Cell Counting Kit-8 CCK-8細(xì)胞計(jì)數(shù)試劑盒的優(yōu)點(diǎn)：

● CCK-8 Kit試劑盒使用方便，省去了洗滌細(xì)胞，不需要放射性同位素和有機(jī)溶劑；
● CCK-8法能快速檢測；
● CCK-8法的檢測靈敏度很高，甚至可以測定較低細(xì)胞密度；
● CCK-8法的重復(fù)性優(yōu)于MTT法；
● CCK-8法對細(xì)胞毒性小；
● CCK-8 Kit細(xì)胞活性檢測試劑中為1瓶溶液，毋需預(yù)制，即開即用。
 

的缺點(diǎn)：

● 與MTT法相比，CCK8和XTT的價(jià)格比較貴；
● CCK8試劑的顏色為淡紅色，與含酚紅的培養(yǎng)基顏色接近，不注意的話容易產(chǎn)生漏加或多加。
 

 主要產(chǎn)品特性及應(yīng)用

★ 操作簡便，省時(shí)省力
★ 水溶性，不需要換液，尤其適合于懸浮細(xì)胞
★ 無需放射性同位素和有機(jī)溶劑，對細(xì)胞毒性低
★ 為1瓶溶液，毋需預(yù)制，即開即用
★ 適合于高通量藥物篩選
 

 的用途：

藥物篩選、細(xì)胞增殖測定、細(xì)胞毒性測定、腫瘤藥敏試驗(yàn)

Storage Condition : 0-5^oC
Shipping Condition : ambient temperature

Product Description
Cell Counting Kit-8 (CCK-8) allows sensitive colorimetric assays for the determination of cell viability in cell proliferation and cytotoxicity assays. Dojindo’s highly water-soluble tetrazolium salt, WST-8*, is reduced by dehydrogenase activities in cells to give a yellow-color formazan dye, which is soluble in the tissue culture media. The amount of the formazan dye, generated by the activities of dehydrogenases in cells, is directly proportional to the number of living cells. The detection sensitivity of CCK-8 is higher than the other tetrazolium salts such as MTT, XTT, MTS or WST-1.
*Patent No. US6,063,587 & CA2,251,850




Cell viability detection mechanism with CCK-8

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Procedure
Cell Counting Kit-8 requires 3 simple steps.


The handling time of Cell Counting Kit-8 is the shortest among it’s competitors. Only 15 minutes of handling time for Cell Counting Kit-8 where as longer handling time is required for both MTS and MTT assay.

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Stability
Cell Counting Kit-8 is a ready-to-use solution. It is also stabe at 4oC for 1 year. Your assay can be done anytime without thaw and freeze.

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Sensitivity
CCK-8 (WST-8) is the highest sensitive dye for the cell based assay.

Comparison of the sensitivity using tetrazolium dye

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Cytotoxicity of Reagents
Only in CCK-8, continious culture is possible without killing cells.

Observation of Cytotoxicity after incubation with each reagent.
HeLa cells were incubated with WST-8 (CCK-8), WST-1, or MTS at 24 hours from addition of those reagents. 

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