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Cell Counting Kit-8 (CCK-8)
TargetMol

为众多的药物研发团队赋能,

让新药发现更简单!

Cell Counting Kit-8 (CCK-8)

产品编号 C0005Cell Counting Kit-8 (CCK-8)
Cell Counting Kit-8 (CCK-8)试剂盒通过采用水溶性四唑盐 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)实现快速检测,其原理是 WST-8在电子耦合试剂存在的情况下,可以被线粒体内的脱氢酶还原生成橙黄色的甲臜染料(formazan)。通过比色,可以动态地量化活细胞的数量,从而对细胞增殖或药物毒性进行检测。CCK-8溶液可直接加入到细胞中,不需要与其它组分预混,操作方便,检测准确。
规格价格库存数量
1 mL (100 T)
¥ 168
现货
5 mL (500 T)
¥ 419
现货
10 mL (1000 T)
¥ 689
现货
30 mL (3000 T)
¥ 1,596
现货
100 mL (10000 T)
¥ 4,760
现货
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TargetMol 的所有产品仅用作科学研究或药证申报,不能被用于人体,我们不向个人提供产品和服务。请您遵守承诺用途,不得违反法律法规规定用于任何其他用途。
实验操作小课堂
Cell Counting Kit-8 (CCK-8)产品问题解答
Cell Counting Kit-8 (CCK-8)水浴加热后,吸光度很低,是不是不能水浴加热?
加完CCK8之后只需要放置在37度恒温培养箱中就可以了,不用水浴加热。吸光度低可能是活细胞数量较少,或者孵育时间较短。
请问 Cell Counting Kit-8 (CCK-8) 加多少到细胞培养液里?
细胞培养液中直接加入 1/10 体积的 Cell Counting Kit-8 (CCK-8),充分混合,保证孔中颜色均一性,但不可有气泡产生。96 孔板,每 100 μL 培养液加入 10 μL 检测试剂。
请问 Cell Counting Kit-8 (CCK-8)外周一圈需要铺满细胞吗?
不建议铺满外周的细胞,外周建议加 PBS 防止内层挥发。
请问下你们公司的CCK8试剂, 对于一般肿瘤细胞来说加后建议孵育多久。吸光值在多少左右正常?
一般是1-4 h,具体时间的话需要根据您的细胞情况和预实验结果来确定。一般吸光值在0.8-1.2左右测定会比较好。
请问CCK8能放-80℃吗?
可以,注意避免反复冻融。
常见问题解答
Cocktail 系列的使用方法?
在室温下解冻,开盖前请低速离心一下,以便将黏附于管壁的液体甩至管底。然后在实验前以 1:100(v/v)稀释至溶液样品(细胞裂解液或组织提取液)。
三款 RIPA 裂解液有什么区别?
RIPA 裂解液(强)、RIPA 裂解液(中)、RIPA 裂解液(弱)的有效裂解成分、裂解强度不同,建议查询官网,有三款RIPA裂解液的比较表。
在做加药实验时,药物对 CCK8 测定是否有影响?如何解决?
有时会有影响。如果药物具有还原性,会和 CCK-8 发生显色反应,增加吸光度。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 450 nm 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。
使用 CCK8 进行实验后,发现读值低,不变色、没差异等,应该怎么办?
CCK8 在孵育 1-4 小时后,OD 读值在 1-2 之间为佳,线性关系比较好。关于 CCK8 的使用问题,通常建议调整细胞状态、增加种板数量、改变加药浓度、延长孵育时间等。
蛋白酶抑制剂 cocktail C0001 使用时加到蛋白液中的有效抑制时间有多久?
这个我们暂时没有准确的数据,因为它对不同样品不同蛋白的保护程度是不一样的。一般单次提取以后,不加蛋白酶抑制剂 cocktail 的样品蛋白会在 24 h 后发生明显降解,加了蛋白酶抑制剂 cocktail 的样品蛋白可以维持 48 h 不降解。如果提取后置于 -20℃ 储存的话,一般加了蛋白酶抑制剂 cocktail 的样品蛋白可以至少储存一个月,冻融三次;但时间长了或者冻融次数多了的话,蛋白还是会发生降解的,不同蛋白降解时间不一样的。
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Handling Instruction | TargetMol 检测原理

Cell Counting Kit-8 (CCK-8)是一种氧化还原反应的指示剂,在电子载体1-Methoxy PMS存在的条件下,利用活细胞中的脱氢酶催化四唑盐WST-8生成甲臜染料,而且甲臜染料的生成量与活细胞的数量成线性关系。

检测方法 MTT法 XTT法 WST-1法 CCK8法
甲臜产物的水溶性
产品性状 粉末 2瓶溶液 溶液 1瓶溶液
使用方法 配成溶液后使用 现配现用 即开即用 即开即用
检测灵敏度 + ++ ++ +++
检测时间 较长 较短 较短 最短
检测波长 560~600nM 420~480nM 420~480nM 430~490nM
细胞毒性 高,细胞形态完全消失 很低,细胞形态不变 很低,细胞形态不变 很低,细胞形态不变
试剂稳定性 一般 较差 一般 很好
批量样品检测 可以 非常适合 非常适合 非常适合
便捷程度 一般 便捷 便捷 非常便捷

Handling Instruction | TargetMol 产品优势

  • CCK-8的检测灵敏度高于其他方法,如MTT,XTT或WST-1;
  • 操作更简单,无需有机溶剂;
  • 对细胞无毒性;
  • 稳定的试剂,即开即用。

Handling Instruction | TargetMol 应用领域

  • 细胞增殖测定
  • 细胞活力检测
  • 细胞因子实验
  • 细胞毒性实验

Handling Instruction | TargetMol 操作说明

1. 接种细胞悬液100μL 于96孔板,在细胞培养箱中(37°C, 5% CO2)孵育。

2. 取出需要检测的细胞,于细胞培养液中直接加入1/10体积的Cell Counting Kit-8 (CCK-8),充分混合,保证孔中颜色均一性,但不可有气泡产生。对96孔板,每 100µl培养液加入10µl检测试剂。

3. 继续于细胞培养箱中培养1-4小时。

4. 酶标仪读数之前,置于孔板于摇床振荡上约1min,确保孔板颜色均匀。

5. 用酶标仪读取450nm光吸收值,计算细胞活性。

细胞存活率=[(实验组-空白组) / (对照组-空白组)] × 100%

细胞抑制率=[(对照组-实验组) / (对照组-空白组)] × 100%

实验组:含细胞、培养基、CCK-8和药物;

对照组:含细胞、培养基、CCK-8,不含药物;

空白组:含培养基、CCK-8,不含细胞、药物。

6. 可选步骤:在100μl细胞中加入10 μl 3 % SDS 以终止反应,可在3天内读取数据。

Handling Instruction | TargetMol 储存条件

4℃避光可保存一年,-20℃避光可保存两年。

Handling Instruction | TargetMol 注意事项

  • 使用前请充分混匀,避免反复冻融
  • 96孔板注意边缘效应,建议弃用周围一圈板孔,改加相同量的PBS。
  • 本产品只能用于研究,严禁用于人体。

SCI 文献

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