上海金畔生物科技有限公司代理UELANDY荧光染料全系系列产品

Cell Counting Kit-8（CCK-8）细胞增殖检测试剂盒

产品货号: C6005S/C6005M/C6005L/C6005XL/C6005XXL

产品规格: 100T/500T/3000T/10000T/5×10000T

目录价（元）:221/516/1886/5158/18421

推荐仪器：酶标仪

大包装询价

产品概述：
储存条件
4℃避光保存，长期储存置于-20℃，有效期见外包装。
  

产品货号	C6005S	C6005M	C6005L	C6005XL	C6005XXL
产品规格	100T	500T	3000T	10000T	5×10000T
组分	1 mL	5 mL	30 mL	100 mL	5×100 mL

 

产品介绍
Cell Counting Kit-8 简称 CCK8（或 WST-8）试剂盒，是一种基于 WST-8（化学名：2-(2-甲氧基-4-硝苯基)-3-(4- 硝苯基)-5-(2,4-二磺基苯)-2H-四唑单钠盐）显色反应的广泛应用于细胞增殖和细胞毒性的快速高灵敏度检测试剂盒。
WST-8 工作原理：在电子耦合试剂存在的条件下，可以被线粒体内的脱氢酶还原生成高度水溶性的橙黄色的甲臜产物（formazan）。颜色的深浅与细胞的增殖成正比，与细胞毒性成反比。使用酶标仪在 450 nm 波长处测定 OD 值，间接反映活细胞数量。
WST-8 是 MTT 的一种升级替代产品，和 MTT 或其它 MTT 类似产品如 XTT、MTS 等相比有明显的优点。首先，MTT 被线粒体内的一些脱氢酶还原生成的甲臜不是水溶性的，需要有特定的溶解液来溶解；而 WST-8、XTT 和 MTS 产生的甲臜都是水溶性的，可以省去后续的溶解步骤。其次，WST-8 产生的甲臜比 XTT 和 MTS 产生的甲臜更易溶解。再次，WST-8 比 XTT 和 MTS 更加稳定，使实验结果更加稳定。另外，WST-8 与 MTT、XTT 相比线性范围更宽，灵敏度更高。
CCK 法广泛应用于药物筛选、细胞增殖测定、细胞毒性测定、肿瘤药敏试验以及生物因子的活性检测等方面。
说明书：

 UE-C6005S/C6005M/C6005L/C6005XL/C6005XXL    
MSDS：
MSDS C6005Cell Counting Kit-8（CCK-8）
客户使用反馈：

常见问题解答：

◆为什么测的吸光度值过低？

1.细胞数量过低，建议增加细胞待测数量；
2.染色难度大，建议增加CCK-8孵育时间

◆数据重复性差？
1.孔板最外圈试剂容易挥发，建议最外一圈的孔只加培养基，不作为测定孔用；
2.CCK-8沾壁，建议加入CCK-8后，轻轻敲击培养板以帮助混匀；
3.细胞数量过多或过少，建议预先在1000-100000个/孔范围内摸索条件；
4.孔内气泡干扰酶标仪检测，检测前需确保每个孔内无气泡

◆在做细胞的加药实验时,药物对CCK-8测定是否有影响?
1.注意如果药物具有还原性(如维生素C),会和CCK-8发生显色反应,增加吸光度；
2.药物中的金属对CCK-8显色有影响:当终浓度为1mM的氯化亚铅、氧化铁、硫酸铜会抑制5%,15%, 90%的显色反应,使灵敏度降级。如果终浓度是10mM的话，将会100%抑制。解决办法:首先要确认药物是否有吸收,在含有药物的培养基中加入CCK-8,测定450nm的吸光度,如果它的吸光度比不含药物的培养基(加CCK-8)的吸光度高,则证明药物有影响,可在加CCK-8之前更换培养基,去掉药物的影响；
3.由于培养基中可能含有氧化还原反应的物质,在正式实验之前,建议使用一个孔作一下检测,有必要先确认培养基和CCK-8是否反应，一般正常的OD值应该在0.4以下

◆可否采用CCK-8法进行药物的IC50检测？
可以,将细胞培养后按照不同浓度的药物处理一定时间后进行CCK-8检测,根据吸光度绘制曲线,计算该药物抑制细胞数量一半时的浓度(IC50)

◆CCK-8对于不同的细胞,灵敏度是否一样？
不一样,悬浮细胞较贴壁细胞难染色。对于贴壁细胞,一般加入CCK-8培养1-4小时吸光度已经很高,但对于悬浮细胞则可能吸光度较低,可以通过延长CCK-8的加入时间或增加细胞数量来解决

◆若是暂不检测OD值，该如何处理?
若暂时不测定 OD 值，可以向每孔中加入 10μL 0.1 M 的 HCl 溶液或者 1% w/v SDS 溶液，并遮盖培养板避光保存在室温条件下。24 小时内测定，吸光度不会发生变化

◆用CCK-8测细胞活力时每次都要做标准曲线吗？
建议每次做，虽然细胞是一样的，但是细胞的状态不一定一样。对于状态不一样的细胞，建议每次做标准曲线

使用本产品的文献：

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参考文献
1.Near-infrared upconversion–activated CRISPR-Cas9 system: A remote-controlled gene editing platform 
应用方向：细胞活力检测：A549细胞