Cell Counting Kit-8 (CCK-8)

mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail

Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.

Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody

Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay

SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.

Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
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我们的产品Cell Counting Kit-8(CCK-8)相比以前的方法为细胞数测定和细胞增殖/毒性检测提供了更方便和灵敏的方法。试剂盒使用了一种高水溶性的四唑盐,WST-8,它在电子耦合试剂存在下还原产生水溶性formazan形式的染料。由脱氢酶产生的formazan的量与活细胞的数量呈直接的线性关系。CCK-8的检测灵敏度高于其他方法,如MTT,XTT,MTS或WST-1等。
CCK-8法的优势
比MTT,MTS或WST-1更敏感
对细胞无毒性
步骤更简单,无需有机溶剂
稳定的试剂,即用型
相关生物数据

相关生物数据

相关生物数据

相关生物数据

相关生物数据

相关生物数据

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相关生物数据

100 tests | 1 mL x 1 |
500 tests | 5 mL x 1 |
1000 tests | 5 mL x 2 |
3000 tests | 5 mL x 6 |
10000 tests | 5 mL x 20 |
CCK-8在0-5°C下,避光保存,储存时间超过一年。如需更长时间储存,请将其储存在-20°C。反复冻融会导致背景增加,干扰您的检测。如果频繁使用,请将试剂盒存放在0-5°C。 |
1)Q:每个孔需要接种的细胞数量是多少?
A:对于贴壁细胞,每孔至少需要1000个细胞(100 μL培养基)。对于白细胞,由于灵敏度较低,每孔至少需要2500个细胞(100 μL培养基)。推荐的96孔板每孔最大细胞数为25000。如果使用24孔或6孔板进行该检测,请计算相应的每孔的细胞数,并调整CCK-8的体积,使其为每孔总液体体积的10%。
2)Q:CCK-8对细胞的毒性大小如何?
A:CCK-8的毒性非常低,在CCK-8测定完成后,相同的细胞可用于其他细胞增殖测定,例如结晶紫测定,中性红测定或DNA荧光测定。(除非细胞极为稀少,不推荐把CCK-8测定过的细胞用于其它实验。)
3)Q:怎么设置空白对照?
A:在不含有细胞的培养基中加入CCK-8,酶标仪检测450 nm的吸光度;若做药物刺激实验,可在不含细胞、含药物的培养基中加入CCK-8。酶标仪检测450 nm的吸光度作为空白对照。
4)Q:CCK-8能否用于细胞染色?
A:CCK-8不能用于细胞染色,CCK-8检测原理是高水溶性的四唑盐WST-8在电子耦合试剂存在下还原产生水溶性formazan形式的染料,WST-8及formazan是高度水溶性的,不会进入细胞内对细胞进行染色。
5)Q:没有450 nm的滤光片,能否使用其他滤光片?
A:如果您没有450 nm滤光片。您也可以使用吸光度在430 nm和490 nm之间的滤光片, 450 nm滤光片具有最佳灵敏度。
6)Q:哪些物质会干扰CCK-8的测定?
A:WST-8可能与还原剂反应生成WST-8 formazan,如果使用还原剂(例如一些抗氧化剂)会增加OD值;若存在氧化性物质则会抑制CCK-8测定反应,减小OD值;此外,培养基中的酚红不会影响实验结果,酚红的吸光度可以在计算时,通过扣除空白孔中本底的吸光度而消去。
7)Q:在做药物刺激实验时,药物对测定是否有影响?
A:如果药物有还原性,则会与CCK-8发生反应,影响吸光度;药物中金属离子的存在可能会影响CCK-8的灵敏度。终浓度为1 mM 的氯化亚铅、氯化铁、硫酸铜会抑制5% 、15% 、90%的显色反应,使灵敏度降低。如果终浓度是10 mM 的话,将会100%抑制。
8)Q:如果吸光度值太低,可以采取什么办法?
A:适当增加细胞数量或延长加入CCK-8溶液后的孵育时间。
9)Q:如果吸光度值太高,可以采取什么办法?
A:适当减少细胞数量(建议正式实验前优化细胞数量与荧光值之间的关系)或缩短加入CCK-8溶液后的孵育时间。