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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):300/700/2560/7000/25000
可替代市场同类产品
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
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
引用及参考文献:
1.Oncogene miR-187-5p is associated with cellular proliferation, migration,invasion, apoptosis and an increased risk of recurrence in bladder cancer
ZuweiLi ,CanbinLin,LiwenZhao,LiangZhou,XiangPan,JingQuan,XiqiPeng,WeiqingLi,HangLi,JinlingXu,WeijieXu,XinGuan,YunChen
Biomedicine & Pharmacotherapy(2018)
应用方向:细胞活性检测:膀胱癌细胞(BC)
2.MicroRNA‑572 functions as an oncogene and a potential biomarker for renal cell carcinoma prognosis
- Xiang Pan, Zuwei Li , Liwen Zhao, Jing Quan, Liang Zhou, Jinling Xu, Weiji Xu, Xin Guan, Hang Li , Shangqi Yang, Yaoting Gui
Oncology Reports(2018) 3092-3101
应用方向:细胞活性检测:肾细胞(RCC)
- 3.Oncogene miR-154-5p regulates cellular function and acts as a molecular marker with poor prognosis in renal cell carcinoma
Lin Canbin, Li Zuwei, Chen Peijie, Quan Jing , Pan Xiang, Zhao Liwen, Zhou Liang, Lai Yulin, He Tao,Xu Weijie,Xu Jinling,Guan Xin,Li Hang, Yang Shangqi,Hu Yimin
Life Sciences(2018)
应用方向:细胞活性检测:膀胱癌细胞(BC)
4.外泌体介导乳腺癌 MCF-7 细胞对多柔比星耐药性的机制
张如月; 李朵璐; 杨哲; 郭金秀; 周玉冰;
药学学报(2019)
应用方向:细胞活性检测:MCF-7、MCF-7/Exo、MCF-7/Dox细胞
5.MS-275联合顺铂对食管鳞状细胞癌9706细胞氧化损伤和凋亡的影响
刘腾飞,王亚苹,李亚,张振坤,李喆,周馨魁,张璐玉,王文杰,刘红涛,张彦婷,关方霞,马珊珊
郑州大学生命科学学院
应用方向:细胞的存活率检测:食管鳞状细胞癌(鳞癌) EC9706细胞
6.A dual-enzymatically cross-linked injectable gelatin hydrogel loaded with BMSC improves neurological function recovery of traumatic brain injury in rats
Minghao Yao, Feng Gao, Ru Xu, Junni Zhang, Yihao Chena , Fangxia Guan
Biomaterials Science
应用方向:细胞活性检测:BMSC
7.重组人MG53蛋白激活Akt/GSK-3β通路降低LPS诱导的hUC-MSCs氧化损伤
王亚苹 张振坤 李喆 刘腾飞 黄团结 周馨魁 麻建杰 关方霞 马珊珊
郑州大学学报(医学版)
应用方向:细胞活力检测:人脐带间充质干细胞(hUC-MSCs)
8.Inonotus Obliquus Polysaccharides Induces Apoptosis of Lung Cancer Cells and Alters Energy Metabolism via the LKB1/AMPK Axis
Shuping Jiang,Fuli Shi,Hui Lin,Ying Ying,Lingyu Luo,Deqiang Huang,Zhijun Luo
International Journal of Biological Macromolecules Volume 151, 15 May 2020, Pages 1277-1286
9.A chitosan-based thermosensitive scaffold loaded with bone marrow-derived mesenchymal stem cells promotes motor function recovery in spinal cord injured mice
Junni Zhang,Tian Cheng,Yihao Chen,Feng Gao,Fangxia Guan,Minghao Yao
Biomedical materials
10.Structure analysis of polysaccharides purified from Cyclocarya paliurus with DEAE-Cellulose and its antioxidant activity in RAW264.7 cells
QiAn,XimeiYe,YiHan,MengZhao,SiChen,XinLiu,XiangLi,ZitongZhao,YangZhang,KehuiOuyang,WenjunWang
International Journal of Biological Macromolecules(Available online 28 November 2019)
11.Dual-enzymatically crosslinked hyaluronic acid hydrogel as a longtime 3D stem cell culture system
Feng Gao,Jinrui Li,Luyu Wang,Dan Zhang,Junni Zhang,Fangxia Guan,Minghao Yao
Biomedical Materials
12.Enhanced effect of nano-monetite hydrosol on dentin remineralization and tubule occlusion
Shenglong Tan,Shangsi Chen,Yifan Wang,Feige Wu,Yufei Shi,Jianglin Wang,Yinyin Du,Shengmin Zhang
Dental Materials(Volume 36, Issue 6, June 2020, Pages 816-825)
13.Control the fate of human umbilical cord mesenchymal stem cells with dual-enzymatically cross-linked gelatin hydrogels for potential applications in nerve regeneration
JinruiLi,FengGao,ShanshanMa,YantingZhang,JunniZhang,FangxiaGuan,MinghaoYao
Journal of Tissue Engineering and Regenerative Medicine
14.MG53 Protects hUC-MSCs against Inflammatory Damage and Synergistically Enhances Their Efficacy in Neuroinflammation Injured Brain through Inhibiting NLRP3/Caspase-1/IL-1β Axis
Shanshan Ma,Yaping Wang,Xinkui Zhou,Zhe Li,Zhenkun Zhang,Yingying Wang,Tuanjie Huang,Yanting Zhang,Jijing Shi,Fangxia Guan
ACS Chem. Neurosci. 2020, 11, 17, 2590–2601
15.Sodium alginate/collagen hydrogel loaded with human umbilical cord mesenchymal stem cells promotes wound healing and skin remodeling
Zhenkun Zhang, Zhe Li, Ya Li, Yingying Wang, Minghao Yao, Kun Zhang, Zhenyu Chen, Han Yue, Jijing Shi, Fangxia Guan & Shanshan Ma
Cell and Tissue Research
16.Reshaping the Immune Microenvironment by Oncolytic Herpes Simplex Virus in Murine Pancreatic Ductal Adenocarcinoma
Liming Zhang,Wei Wang,Ruikun Wang,Jie Yin,Hongkai Zhang,Youjia Cao
Molecular Therapy(VOLUME 29, ISSUE 2, P744-761, FEBRUARY 03, 2021)
17.Selective extracellular arginine deprivation by a single injection of cellular non-uptake arginine deiminase nanocapsules for sustained tumor inhibition
Hongzhao Qi,Yin Wang,Xubo Yuan,Peifeng Lia,Lijun Yang
Nanoscale
18.Inducing Apoptosis and Suppressing Inflammatory Reactions in Synovial Fibroblasts are Two Important Ways for Guizhi-Shaoyao-Zhimu Decoction Against Rheumatoid Arthritis
Zhang Q, Duan HX, Li RL, Sun JY, Liu J, Peng W, Wu CJ, Gao YX
Journal of Inflammation Research(26 January 2021 Volume 2021:14 Pages 217—236)
19.Injectable bone cement with magnesium-containing microspheres enhances osteogenesis via anti-inflammatory immunoregulation
Shenglong Tan,Yifan Wang,Yingying Du,Yin Xiao,Shengmin Zhang
Bioactive Materials Volume 6, Issue 10, October 2021, Pages 3411-3423
20.Propionate and butyrate produced by gut microbiota after probiotic supplementation attenuate lung metastasis of melanoma cells in mice
Lili Chen,Xinyu Zhou,Yawei Wang,Dake Wang,Yueshuang Ke,Xianlu Zeng
Molecular Nutrition & Food Research
21.Regulating the Anticancer Efficacy of Sgc8-Combretastatin A4 Conjugates: A Case of Recognizing the Significance of Linker Chemistry for the Design ofAptamer-Based Targeted Drug Delivery Strategies
Zhiyong Huang,Dan Wang,Cheng-Yu Long,Shen-Huan Li,Xue-Qiang Wang,Weihong Tan
Journal of American Chemical Society
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
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