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Cell Counting Kit-8（CCK-8）細(xì)胞增殖檢測(cè)試劑盒
產(chǎn)品貨號(hào): C6005S/C6005M/C6005L/C6005XL/C6005XXL
產(chǎn)品規(guī)格: 100T/500T/3000T/10000T/5×10000T
目錄價(jià)（元）:300/700/2560/7000/25000
可替代市場(chǎng)同類(lèi)產(chǎn)品
產(chǎn)品概述：
儲(chǔ)存條件
4℃避光保存，長(zhǎng)期儲(chǔ)存置于-20℃，有效期見(jiàn)外包裝。
產(chǎn)品介紹
Cell Counting Kit-8 簡(jiǎn)稱(chēng) CCK8（或 WST-8）試劑盒，是一種基于 WST-8（化學(xué)名：2-(2-甲氧基-4-硝苯基)-3-(4- 硝苯基)-5-(2,4-二磺基苯)-2H-四唑單鈉鹽）顯色反應(yīng)的廣泛應(yīng)用于細(xì)胞增殖和細(xì)胞毒性的快速高靈敏度檢測(cè)試劑盒。
WST-8 工作原理：在電子耦合試劑存在的條件下，可以被線(xiàn)粒體內(nèi)的脫氫酶還原生成高度水溶性的橙黃色的甲臜產(chǎn)物（formazan）。顏色的深淺與細(xì)胞的增殖成正比，與細(xì)胞毒性成反比。使用酶標(biāo)儀在 450 nm 波長(zhǎng)處測(cè)定 OD 值，間接反映活細(xì)胞數(shù)量。
WST-8 是 MTT 的一種升級(jí)替代產(chǎn)品，和 MTT 或其它 MTT 類(lèi)似產(chǎn)品如 XTT、MTS 等相比有明顯的優(yōu)點(diǎn)。首先，MTT 被線(xiàn)粒體內(nèi)的一些脫氫酶還原生成的甲臜不是水溶性的，需要有特定的溶解液來(lái)溶解；而 WST-8、XTT 和 MTS 產(chǎn)生的甲臜都是水溶性的，可以省去后續(xù)的溶解步驟。其次，WST-8 產(chǎn)生的甲臜比 XTT 和 MTS 產(chǎn)生的甲臜更易溶解。再次，WST-8 比 XTT 和 MTS 更加穩(wěn)定，使實(shí)驗(yàn)結(jié)果更加穩(wěn)定。另外，WST-8 與 MTT、XTT 相比線(xiàn)性范圍更寬，靈敏度更高。
CCK 法廣泛應(yīng)用于藥物篩選、細(xì)胞增殖測(cè)定、細(xì)胞毒性測(cè)定、腫瘤藥敏試驗(yàn)以及生物因子的活性檢測(cè)等方面。
說(shuō)明書(shū)：
UE-C6005S/C6005M/C6005L/C6005XL/C6005XXL
客戶(hù)使用反饋：
常見(jiàn)問(wèn)題解答：
◆為什么測(cè)的吸光度值過(guò)低？
1.細(xì)胞數(shù)量過(guò)低，建議增加細(xì)胞待測(cè)數(shù)量；
2.染色難度大，建議增加CCK-8孵育時(shí)間
◆數(shù)據(jù)重復(fù)性差？
1.孔板最外圈試劑容易揮發(fā)，建議最外一圈的孔只加培養(yǎng)基，不作為測(cè)定孔用；
2.CCK-8沾壁，建議加入CCK-8后，輕輕敲擊培養(yǎng)板以幫助混勻；
3.細(xì)胞數(shù)量過(guò)多或過(guò)少，建議預(yù)先在1000-100000個(gè)/孔范圍內(nèi)摸索條件；
4.孔內(nèi)氣泡干擾酶標(biāo)儀檢測(cè)，檢測(cè)前需確保每個(gè)孔內(nèi)無(wú)氣泡
◆在做細(xì)胞的加藥實(shí)驗(yàn)時(shí),藥物對(duì)CCK-8測(cè)定是否有影響?
1.注意如果藥物具有還原性(如維生素C),會(huì)和CCK-8發(fā)生顯色反應(yīng),增加吸光度；
2.藥物中的金屬對(duì)CCK-8顯色有影響:當(dāng)終濃度為1mM的氯化亞鉛、氧化鐵、硫酸銅會(huì)抑制5%,15%, 90%的顯色反應(yīng),使靈敏度降級(jí)。如果終濃度是10mM的話(huà)，將會(huì)100%抑制。解決辦法:首先要確認(rèn)藥物是否有吸收,在含有藥物的培養(yǎng)基中加入CCK-8,測(cè)定450nm的吸光度,如果它的吸光度比不含藥物的培養(yǎng)基(加CCK-8)的吸光度高,則證明藥物有影響,可在加CCK-8之前更換培養(yǎng)基,去掉藥物的影響；
3.由于培養(yǎng)基中可能含有氧化還原反應(yīng)的物質(zhì),在正式實(shí)驗(yàn)之前,建議使用一個(gè)孔作一下檢測(cè),有必要先確認(rèn)培養(yǎng)基和CCK-8是否反應(yīng)，一般正常的OD值應(yīng)該在0.4以下
◆可否采用CCK-8法進(jìn)行藥物的IC50檢測(cè)？
可以,將細(xì)胞培養(yǎng)后按照不同濃度的藥物處理一定時(shí)間后進(jìn)行CCK-8檢測(cè),根據(jù)吸光度繪制曲線(xiàn),計(jì)算該藥物抑制細(xì)胞數(shù)量一半時(shí)的濃度(IC50)
◆CCK-8對(duì)于不同的細(xì)胞,靈敏度是否一樣？
不一樣,懸浮細(xì)胞較貼壁細(xì)胞難染色。對(duì)于貼壁細(xì)胞,一般加入CCK-8培養(yǎng)1-4小時(shí)吸光度已經(jīng)很高,但對(duì)于懸浮細(xì)胞則可能吸光度較低,可以通過(guò)延長(zhǎng)CCK-8的加入時(shí)間或增加細(xì)胞數(shù)量來(lái)解決
◆若是暫不檢測(cè)OD值，該如何處理?
若暫時(shí)不測(cè)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，并遮蓋培養(yǎng)板避光保存在室溫條件下。24 小時(shí)內(nèi)測(cè)定，吸光度不會(huì)發(fā)生變化
◆用CCK-8測(cè)細(xì)胞活力時(shí)每次都要做標(biāo)準(zhǔn)曲線(xiàn)嗎？
建議每次做，雖然細(xì)胞是一樣的，但是細(xì)胞的狀態(tài)不一定一樣。對(duì)于狀態(tài)不一樣的細(xì)胞，建議每次做標(biāo)準(zhǔn)曲線(xiàn)
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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)
應(yīng)用方向：細(xì)胞活性檢測(cè)：膀胱癌細(xì)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
應(yīng)用方向：細(xì)胞活性檢測(cè)：腎細(xì)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)
應(yīng)用方向：細(xì)胞活性檢測(cè)：膀胱癌細(xì)胞（BC）
4.外泌體介導(dǎo)乳腺癌 MCF-7 細(xì)胞對(duì)多柔比星耐藥性的機(jī)制
張如月； 李朵璐； 楊哲； 郭金秀； 周玉冰；
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應(yīng)用方向：細(xì)胞活性檢測(cè)：MCF-7、MCF-7/Exo、MCF-7/Dox細(xì)胞
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應(yīng)用方向：細(xì)胞的存活率檢測(cè)：食管鱗狀細(xì)胞癌(鱗癌) EC9706細(xì)胞
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Minghao Yao, Feng Gao, Ru Xu, Junni Zhang, Yihao Chena , Fangxia Guan
Biomaterials Science
應(yīng)用方向：細(xì)胞活性檢測(cè)：BMSC
7.重組人MG53蛋白激活A(yù)kt/GSK-3β通路降低LPS誘導(dǎo)的hUC-MSCs氧化損傷
王亞蘋(píng) 張振坤 李喆 劉騰飛 黃團(tuán)結(jié) 周馨魁 麻建杰 關(guān)方霞 馬珊珊
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應(yīng)用方向：細(xì)胞活力檢測(cè)：人臍帶間充質(zhì)干細(xì)胞(hUC-MSCs)
8.Inonotus Obliquus Polysaccharides Induces Apoptosis of Lung Cancer Cells and Alters Energy Metabolism via the LKB1/AMPK Axis
Shuping Jiang，F(xiàn)uli Shi，Hui Lin，Ying Ying，Lingyu Luo，Deqiang Huang，Zhijun Luo
International Journal of Biological Macromolecules Volume 151, 15 May 2020, Pages 1277-1286
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Biomedical materials
10.Structure analysis of polysaccharides purified from Cyclocarya paliurus with DEAE-Cellulose and its antioxidant activity in RAW264.7 cells
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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，F(xiàn)eige 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，F(xiàn)engGao，ShanshanMa，YantingZhang，JunniZhang，F(xiàn)angxiaGuan，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
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16.Reshaping the Immune Microenvironment by Oncolytic Herpes Simplex Virus in Murine Pancreatic Ductal Adenocarcinoma
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17.Selective extracellular arginine deprivation by a single injection of cellular non-uptake arginine deiminase nanocapsules for sustained tumor inhibition
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18.Inducing Apoptosis and Suppressing Inflammatory Reactions in Synovial Fibroblasts are Two Important Ways for Guizhi-Shaoyao-Zhimu Decoction Against Rheumatoid Arthritis
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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
參考文獻(xiàn)
1.Near-infrared upconversion–activated CRISPR-Cas9 system: A remote-controlled gene editing platform
應(yīng)用方向：細(xì)胞活力檢測(cè)：A549細(xì)胞
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